Transformant and method for producing resorcinol using the same

Abstract

To provide a transformant capable of biologically producing resorcinol using saccharide as a raw material.SOLUTION: In one aspect, the present invention relates to a transformant obtained by expressibly introducing (A) genes encoding enzymes having an activity of hydroxylating salicylic acids to produce 2,4-dihydroxy benzoic acids and (B) genes encoding enzymes having an activity of decarboxylating 2,4-dihydroxy benzoic acids into microbial hosts. In another aspect, the present invention relates to a transformant obtained by expressibly introducing cyp genes encoding cytochrome P450, cpr genes encoding cytochrome P450 reductase, and at least one selected from the group consisting of ptaG genes, graF genes, sdc genes, and ubiD genes encoding decarboxylase into a microbial host.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] Resorcin (CAS number: 108-46-3; synonyms include resorcinol and 1,3-dihydroxybenzene) has a wide range of applications in various industries, including rubber, automobiles, chemicals, and construction. It is a necessary raw material for many chemical products, such as rubber and tire adhesives, wood product adhesives, flame retardants, UV absorbers, polycarbonates, pesticides, urethane elastomers, and dyes.

[0002] Traditionally, resorcinol has been produced chemically from petroleum-derived raw materials (such as m-diisopropylbenzene and benzenemethane disulfonic acid). In recent years, in order to reduce carbon dioxide emissions, there has been a demand to produce resorcinol from renewable raw materials, such as non-edible biomass-derived raw materials, instead of petroleum-derived raw materials.

[0003] Meanwhile, the technology of producing salicylic acid from sugar raw materials via chorismate is disclosed in the following documents, for example. Patent Document 1 discloses a technology of "introducing one or more genes encoding enzymes that enable the microorganism to synthesize aromatic compounds from chorismate or isochorismate into a microorganism" in the "method of producing a microorganism with a modified sugar metabolic pathway", or a technology of "introducing a gene encoding isochorismate synthase and a gene encoding isochorismate pyruvate lyase, in which the aromatic compound is salicylic acid". Patent Document 2 discloses a technology (a so-called bifunctional enzyme technology) that "can improve salicylic acid productivity by exogenously expressing a fusion protein in which PchB is bound to the carboxy terminus of MenF". Non-Patent Document 1 discloses a technology of producing salicylic acid from chorismate using "salicylate synthase" derived from "Amycolatopsis methanolica". [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. 2017 / 033965 (claims 1, 3, figure 7, etc.) [Patent Document 2] JP 2016-86688 A (Abstract, etc.) [Non-patent literature]

[0005] [Non-Patent Document 1] Appl Microbiol Biotechnol 99(14):5895-905(2015). Summary of the Invention [Problem to be solved by the invention]

[0006] The present disclosure provides a transformant capable of biologically producing resorcinol using sugars and the like as raw materials, and a method for producing resorcinol using the transformant. [Means for solving the problem]

[0007] In one aspect, the present disclosure relates to a transformant obtained by expressibly introducing into a microbial host a gene (A) encoding an enzyme having an activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid and a gene (B) encoding an enzyme having 2,4-dihydroxybenzoic acid decarboxylation activity.

[0008] In another aspect, the present disclosure relates to a transformant obtained by expressibly introducing into a microbial host a cyp gene encoding a cytochrome P450, a cpr gene encoding a cytochrome P450 reductase, and at least one gene selected from the group consisting of a ptaG gene, a graF gene, an sdc gene, and an ubiD gene encoding a decarboxylase.

[0009] In another aspect, the present disclosure relates to a method for producing resorcinol, comprising a step of culturing the transformant of the present disclosure in the presence of sugars to produce resorcinol. Effect of the Invention

[0010] According to one aspect of the present disclosure, a transformant capable of producing resorcinol using saccharides or the like as raw materials can be provided. According to another aspect of the present disclosure, resorcinol can be biologically produced (produced) using the transformant. [Brief description of the drawings]

[0011] [Figure 1] FIG. 1 illustrates a schematic diagram of resorcin production in the transformant of the present disclosure. [Diagram 2] FIG. 2 is an overall metabolic pathway diagram illustrating the biosynthetic pathway of resorcin in one embodiment of the transformant of the present disclosure. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] The present disclosure is based on the discovery by the present inventors that a transformant capable of biologically producing resorcinol using sugars as raw materials can be obtained by expressing a gene (A) encoding an enzyme having the activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid and a gene (B) encoding an enzyme having 2,4-dihydroxybenzoic acid decarboxylation activity into a microbial host. The present disclosure is also based on the discovery by the present inventors that a transformant capable of biologically producing resorcinol using sugars as raw materials can be obtained by introducing into a microbial host a cyp gene encoding cytochrome P450, a cpr gene encoding cytochrome P450 reductase (abbreviated as CYP reductase, EC number: EC 1.6.2.4), and a ptaG gene, a graF gene, a sdc gene, or a ubiD gene encoding a decarboxylase, in an expressible manner.

[0013] In the present disclosure, in one or more embodiments, "introduction of a gene" refers to introduction of the gene so that the gene can be expressed in a host. In one or more embodiments, gene introduction into a host can be performed using a general gene recombination technique (for example, the method described in Michael R. Green & Joseph Sambrook, Molecular cloning, Cold Spring Harbor Laboratory Press). In one or more embodiments, examples of gene introduction include gene introduction using a plasmid vector, or incorporation into the chromosome of a microbial host.

[0014] In the present disclosure, "85% or more identity" with respect to an amino acid sequence or a nucleotide sequence means having at least 85% identity, and in one or more embodiments, refers to identity of 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more.

[0015] In the present disclosure, "identity of amino acid sequence or nucleotide sequence" can be performed using a readily available sequence comparison computer program. In one or more embodiments, the computer program may include the GCG Wisconsin Bestfit package (University of Wisconsin, USA; Devereux et al. (1984) Nucleic Acids Res. 12: 387), the BLAST package (Ausubel et al. (1999) ibid-Ch. 18), and FASTA (Atschul et al. (1990) J. Mol. Biol. 403-410).

[0016] In the present disclosure, "stringent conditions" refers to conditions under which so-called specific hybrids are formed and non-specific hybrids are not formed. In one or more embodiments, stringent conditions include conditions under which highly identical base sequences hybridize with each other, but base sequences with lower identity do not hybridize with each other. In one or more embodiments, high identity between base sequences includes identity of 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, or 97% or more. In one or more embodiments, stringent conditions may be conditions described in Molecular Cloning, A Laboratory Manual, Second Edition, 1989, Vol2, p11.45. Specifically, hybridization may occur at a temperature 5°C to 10°C lower than the melting temperature (Tm) of a complete hybrid.

[0017] [Microbial host] The microbial host according to the present disclosure is not particularly limited, and may be, for example, a coryneform bacterium, Escherichia coli (bacteria of the genus Escherichia, particularly Escherichia coli), a solvent-tolerant bacterium, or a yeast. Coryneform bacteria are a group of microorganisms defined in Bergey's Manual of Determinative Bacteriology, Vol. 8, 599 (1974), and are not particularly limited as long as they grow under normal aerobic conditions. In one or more embodiments, the coryneform bacteria include bacteria of the genus Corynebacterium, Brevibacterium, Arthrobacter, Mycobacterium, and Micrococcus. In one or a plurality of embodiments, examples of the solvent-resistant bacteria include Pseudomonas putida S12, Pseudomonas aeruginosa, Pseudomonas paucimobilis, Pseudomonas alcaligenes, Pseudomonas fluorescens, Pseudomonas fragi, Pseudomonas oleovorans, Pseudomonas sp., Rhodococcus erythropolis, Rhodococcus opacus, Burkholderia cepacia, and Paenibacillus illinoisensis.

[0018] The microbial host according to the present disclosure is not particularly limited, and may be, for example, a bacterium of the genus Corynebacterium from the viewpoint of improving resorcin productivity. In one or more embodiments, examples of the bacterium of the genus Corynebacterium include Corynebacterium glutamicum, Corynebacterium efficiens, Corynebacterium ammoniagenes, Corynebacterium halotolerance, and Corynebacterium alkanolyticum. The microbial host according to the present disclosure is not particularly limited, and may be, for example, Corynebacterium glutamicum from the viewpoint of improving resorcin productivity.

[0019] The microbial host according to the present disclosure is not particularly limited, and may be, for example, Corynebacterium glutamicum R (FERM BP-18976), ATCC13032, or ATCC13869, from the viewpoint of improving resorcin productivity.

[0020] In one or more embodiments, the microbial host according to the present disclosure may be a microbial host capable of producing salicylic acid. In the present disclosure, the term "microbial host capable of producing salicylic acid" refers to a microbial host capable of expressing at least an enzyme having salicylic acid synthase activity. The microbial host according to the present disclosure may be not only a bacterium having a gene encoding an enzyme having salicylic acid synthase activity as a wild type, but also a bacterium artificially transformed so as to be capable of expressing an enzyme having salicylic acid synthase activity. In one or more embodiments, the microbial host capable of producing salicylic acid may be a bacterium obtained by transforming a bacterium having a gene encoding an enzyme having salicylic acid synthase activity as a wild type in a state capable of expressing an enzyme having salicylic acid synthase activity.

[0021] In one or more embodiments, the microbial host capable of producing salicylic acid is preferably a bacterium into which a gene encoding an enzyme having salicylic acid synthase activity has been introduced by genetic modification in order to improve resorcinol productivity. The enzyme having salicylic acid synthase activity refers to an enzyme having an activity of catalyzing the reaction of synthesizing salicylic acid from chorismic acid. Whether or not the enzyme having salicylic acid synthase activity is expressed can be confirmed, for example, by adding the bacterial cell or cell disruption solution to be measured to a reaction solution containing chorismic acid, carrying out a reaction of synthesizing salicylic acid, and analyzing the produced salicylic acid by an appropriate method such as HPLC. In one or more embodiments, the microbial host according to the present disclosure may be a microbial host in which the expression level of an enzyme having salicylic acid synthase activity is improved from the viewpoint of improving resorcin productivity, a microbial host that overexpresses an enzyme having salicylic acid synthase activity or is capable of improving the expression level or inducing the overexpression, etc. The expression level of the enzyme can be appropriately adjusted or improved by a person skilled in the art by using a suitable promoter, for example.

[0022] In one or more embodiments, the gene encoding the enzyme having salicylic acid synthase activity may be an irp gene, and from the viewpoint of improving resorcin productivity, the irp gene derived from Amycolatopsis methanolica is preferable. In one or more embodiments, the irp gene derived from Amycolatopsis methanolica may be a gene consisting of the base sequence shown in SEQ ID NO: 64. In one or more embodiments, the gene encoding the enzyme having salicylic acid synthase activity may be a gene including a base sequence having an identity of 85% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, 99.9% or more, or 100% to SEQ ID NO: 64, or a gene consisting of the base sequence.

[0023] [Transformants] In one aspect, the transformant according to the present disclosure is a transformant obtainable by expressibly introducing the above-mentioned gene (A) and gene (B) into a microbial host. In another aspect, the transformant according to the present disclosure is a transformant obtainable by expressibly introducing a cyp gene, a cpr gene, and at least one gene selected from the group consisting of a ubiD gene, a ptaG gene, a graF gene, and an sdc gene into a microbial host. In one or more embodiments, the transformant according to the present disclosure has resorcin-producing ability and can therefore be used for resorcin production. As shown in Fig. 1, in one or more embodiments, the transformant according to the present disclosure can produce salicylic acid from sugars in the living body of the transformant, and can produce resorcin from salicylic acid by gene (A) and gene (B). In one or more embodiments, the cyp gene can be used as the gene (A) to be introduced, and it is preferable to use the cyp gene and the cpr gene from the viewpoint of improving the productivity of resorcin. In one or more embodiments, the ubiD gene, ptaG gene, graF gene, and / or sdc gene can be used as the gene (B) to be introduced, and it is preferable to use the ptaG gene and / or the graF gene from the viewpoint of improving the productivity of resorcin.

[0024] [Gene (A)] In the present disclosure, the gene (A) may be one type of gene or a combination of two or more types of genes in one or more embodiments. In one or more embodiments, the gene (A) encoding an enzyme having an activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid may be, but is not limited to, a gene cyp encoding cytochrome P450, and from the viewpoint of improving resorcinol productivity, it is preferable to include at least a cyp gene and a gene cpr encoding cytochrome P450 reductase (abbreviated as CYP reductase; EC number: EC 1.6.2.4). In one or more embodiments, a polypeptide encoded by the cyp gene (e.g., cytochrome P450, etc.) may have an activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid in combination with a polypeptide encoded by the cpr gene (e.g., cytochrome P450 reductase, etc.).

[0025] In one or more embodiments, from the viewpoint of improving resorcin productivity, it is also preferable to express the polypeptide encoded by the cyp gene in cells together with the polypeptide encoded by the cpr gene. Most of the polypeptides encoded by the cyp gene are polypeptides consisting of about 500 amino acid residues, and are a type of hemoprotein having heme at the active site. The polypeptide encoded by the cyp gene supplies oxygen atoms to the reaction substrate by utilizing oxygen and electrons transferred from iron in the active center and nicotinamide adenine dinucleotide phosphate (NADPH) via the electron transport system. The electron transport system here refers to the polypeptide encoded by the cpr gene.

[0026] The polypeptide encoded by the cpr gene is known as a protein that requires two flavin molecules, namely flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN), as cofactors. The polypeptide encoded by the cpr gene generally has a membrane-binding region, an FAD domain, and an FMN domain, and transfers electrons donated by NADPH to a heme protein via FAD and FMN in order. In this case, the target to which electrons are transferred is a polypeptide encoded by the cyp gene.

[0027] Polypeptides encoded by cyp genes have high substrate specificity and may exist in multiple types within a cell. Polypeptides encoded by cpr genes are known to function as targets for polypeptides encoded by various types of cyp genes.

[0028] The origin of the cyp gene and the cpr gene is as described below. In one or more embodiments, the cyp gene and the cpr gene may be introduced in combination from the same genus (or species), but it is not necessary to introduce the cyp gene and the cpr gene from the same genus (or species) in combination. Even when the cyp gene and the cpr gene from any different genera (or species) are introduced in combination into the host, the polypeptide encoded by the cyp gene may have the activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid in combination with the polypeptide encoded by the cpr gene. The names (designations) of the "genus" and "species" of the organisms from which the genes are derived shown in the present disclosure are merely examples, and there may be synonymous alternative names.

[0029] In one or more embodiments, whether or not the gene is a "gene encoding an enzyme having the activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid" can be evaluated by carrying out a reaction of producing 2,4-dihydroxybenzoic acid using salicylic acid as a substrate, using a transformant prepared by expressibly introducing the gene to be evaluated into Corynebacterium glutamicum. When the production of 2,4-dihydroxybenzoic acid or an increase in the amount of production is confirmed, it can be determined that the gene is a gene encoding an enzyme having the activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid. The production of 2,4-dihydroxybenzoic acid can be measured based on the description of the examples in the present specification. In the present disclosure, the "activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid" may also be referred to as hydroxylase activity against salicylic acid.

[0030] In one or more embodiments, the enzyme encoded by gene (A) may have an activity of hydroxylating benzoic acid to produce 4-hydroxybenzoic acid (also referred to as hydroxylase activity for benzoic acid). In one or more embodiments, whether or not the enzyme has hydroxylase activity for benzoic acid can be evaluated by carrying out a reaction of hydroxylating benzoic acid to produce 4-hydroxybenzoic acid using a transformant prepared by introducing the gene to be evaluated into Corynebacterium glutamicum so that the gene can be expressed. When production of 4-hydroxybenzoic acid or an increase in the amount of production is confirmed, it can be determined that the enzyme has the activity of hydroxylating benzoic acid to produce 4-hydroxybenzoic acid. The production of 4-hydroxybenzoic acid can be measured based on the description in the Examples of the present specification. In one or more embodiments, the enzyme encoded by gene (A) may have a hydroxylase activity for benzoic acid that is substantially the same as, higher than, or lower than the hydroxylase activity for salicylic acid. In one or more embodiments, the hydroxylase activity for benzoic acid being substantially the same as the hydroxylase activity for salicylic acid may be 85% to 120% or 90% to 120% of the hydroxylase activity for salicylic acid. In one or more embodiments, the hydroxylase activity for benzoic acid being higher than the hydroxylase activity for salicylic acid may be 200% or more or 300% or more of the hydroxylase activity for salicylic acid. In one or more embodiments, the hydroxylase activity for benzoic acid being lower than the hydroxylase activity for salicylic acid may mean that the hydroxylase activity for benzoic acid is 50% or less or 45% or less of the hydroxylase activity for salicylic acid. In one or more embodiments, the comparison of the hydroxylase activity for benzoic acid and the hydroxylase activity for salicylic acid can be performed by carrying out the hydroxylase reaction using a transformant prepared by introducing the gene to be evaluated into Corynebacterium glutamicum so that it can be expressed, under the same culture conditions except for the substrate being different (benzoic acid or salicylic acid), and comparing the products produced.

[0031] [cyp gene] The origin of the cyp gene is not particularly limited, and examples thereof include the genera Aspergillus, Ustilago, Phanerochaete, Cochliobolus, Neurospora, and Rhodotorula, etc. In one or more embodiments, the origin of the cyp gene is preferably the genera Aspergillus, Phanerochaete, Cochliobolus, Neurospora, and Rhodotorula, and more preferably the genera Aspergillus and Rhodotorula, from the viewpoint of improving resorcin productivity.

[0032] The genus Aspergillus according to the present disclosure is not particularly limited, for example, Aspergillus niger, Aspergillus clavatus, Aspergillus oryzae, Aspergillus fumigatus, Aspergillus phoencis, Aspergillus tubingensis, Aspergillus luchuensis, Aspergillus brasiliensis, Aspergillus piperis, Aspergillus costaricaensis, Aspergillus eucalypticola, Aspergillus vadensis, Aspergillus welwitschiae, Aspergillus ibericus, Aspergillus sclerotioniger, Aspergillus carbonarius, Aspergillus sclerotiicarbonarius, Aspergillus homomorphus, Aspergillus brunneoviolaceus, Aspergillus fijiensis, Aspergillus japonicus, Aspergillus uvarum, Aspergillus nomiae, Aspergillus parasiticus, Aspergillus tamarii, Aspergillus turcosus, Aspergillus caelatus, Aspergillus pseudotamarii, Aspergillus lentulus, Aspergillus udagawae, Aspergillus viridinutans, Aspergillus pseudoviridinutans, Aspergillus fumigatiaffinis, Aspergillus ochraceoroseus, Aspergillus wentii, Aspergillus melleus, Aspergillus hiratsukae, Aspergillus felis, Aspergillus rambellii, Aspergillus arachidicola, Aspergillus thermomutatus, AspergillusAspergillus fischeri, Aspergillus terreus, Aspergillus puulaauensis, Aspergillus nidulans, Aspergillus calidoustus, and Aspergillus tanneri, and may be Aspergillus niger or Aspergillus clavatus. Examples of the cyp gene derived from Aspergillus niger include a gene consisting of the base sequence shown in SEQ ID NO:1, and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:69.

[0033] The genus Ustilago according to the present disclosure is not particularly limited, and examples thereof include Ustilago maydis, Ustilago loliicola, Ustilago hordei, Ustilago nuda, Ustilago bromivora, Ustilago trichophora, and Ustilago tritici, and may be Ustilago maydis. Examples of the cyp gene derived from Ustilago maydis include a gene consisting of the nucleotide sequence shown in SEQ ID NO:4, and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:70.

[0034] The genus Phanerochaete according to the present disclosure is not particularly limited, and may be, for example, Phanerochaete chrysosporium, Phanerochaete sordida, and Phanerochaete carnosa, and may be Phanerochaete chrysosporium. The genus Phanerochaete is also synonymous with the genus Phanerodontia, and Phanerochaete chrysosporium, Phanerochaete sordida, and Phanerochaete carnosa may be, respectively, Phanerodontia chrysosporium, Phanerodontia sordida, and Phanerodontia carnosa. Examples of the cyp gene derived from Phanerochaete chrysosporium (Phanerodontia chrysosporium) include a gene consisting of the nucleotide sequence shown in SEQ ID NO: 7, and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 71.

[0035] The genus Cochliobolus according to the present disclosure is not particularly limited, and examples thereof include Cochliobolus lunatus, Cochliobolus victoriae, Cochliobolus heterostrophus, Cochliobolus carbonum, Cochliobolus sativus, Cochliobolus miyabeanus, Cochliobolus kusanoi, and Cochliobolus spicifer, and may be Cochliobolus lunatus. Examples of the cyp gene derived from Cochliobolus lunatus include a gene consisting of the base sequence shown in SEQ ID NO: 10, and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 72.

[0036] The genus Neurospora according to the present disclosure is not particularly limited, and examples thereof include Neurospora crassa, Neurospora tetrasperm, Neurospora discreta, Neurospora intermedia, Neurospora sitophila, and Neurospora metzenbergii, and may be Neurospora crassa. Examples of the cyp gene derived from Neurospora crassa include a gene consisting of the nucleotide sequence shown in SEQ ID NO:13, and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:73.

[0037] The genus Rhodotorula according to the present disclosure is not particularly limited, and examples thereof include Rhodotorula minuta, Rhodotorula gracilis, Rhodotorula rubescens, Rhodotorula creatinivora, Rhodotorula diobovata, Rhodotorula bogoriensis, and Rhodotorula mucilaginosa, and may be Rhodotorula minuta. Examples of the cyp gene derived from Rhodotorula minuta include a gene consisting of the nucleotide sequence shown in SEQ ID NO:16 and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:74.

[0038] The cyp gene is not particularly limited and may be, for example, any one of the following genes (A1), (A2), and (A3). (A1) A gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 69, 70, 71, 72, 73, or 74, or a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 69, 70, 71, 72, 73, or 74. (A2) A gene encoding a polypeptide having an amino acid sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 69, 70, 71, 72, 73, or 74, and which encodes a polypeptide having an activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid. or a gene encoding a polypeptide consisting of an amino acid sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 69, 70, 71, 72, 73 or 74, and which encodes a polypeptide having the activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid. (A3) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 69, 70, 71, 72, 73 or 74, and that encodes a polypeptide having the activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid, or a gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 69, 70, 71, 72, 73 or 74, and that encodes a polypeptide having the activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid. In one or more embodiments, the polypeptides encoded by the genes (A2) and (A3), in combination with the polypeptide encoded by the cpr gene, may exhibit activity to hydroxylate salicylic acid to produce 2,4-dihydroxybenzoic acid.

[0039] The cyp gene is not particularly limited and may be, for example, any one of the following genes (A4), (A5), and (A6). (A4) A gene having the base sequence shown in SEQ ID NO: 1, 4, 7, 10, 13 or 16, or a gene consisting of the base sequence shown in SEQ ID NO: 1, 4, 7, 10, 13 or 16. (A5) A gene having a nucleotide sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to a gene having a nucleotide sequence shown in SEQ ID NO: 1, 4, 7, 10, 13, or 16, and encoding a polypeptide having an activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid. or a gene having a base sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the base sequence shown in SEQ ID NO: 1, 4, 7, 10, 13 or 16, and encoding a polypeptide having the activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid. (A6) A gene that hybridizes under stringent conditions with a gene having a nucleotide sequence complementary to a gene having the nucleotide sequence shown in SEQ ID NO: 1, 4, 7, 10, 13 or 16 and encodes a polypeptide having the activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid, or a gene that hybridizes under stringent conditions with a gene having a nucleotide sequence complementary to a gene consisting of the nucleotide sequence shown in SEQ ID NO: 1, 4, 7, 10, 13 or 16 and encodes a polypeptide having the activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid. In one or more embodiments, the polypeptides encoded by the genes (A5) and (A6), in combination with the polypeptide encoded by the cpr gene, may exhibit activity to hydroxylate salicylic acid to produce 2,4-dihydroxybenzoic acid.

[0040] In one or more embodiments, the cyp gene is preferably a cyp gene derived from Aspergillus niger, a cyp gene derived from Phanerochaete chrysosporium, or a cyp gene derived from Rhodotorula minuta from the viewpoint of improving resorcin productivity. From the same viewpoint, in one or more embodiments, the cyp gene is preferably a gene having a base sequence shown in SEQ ID NO: 1, 7, or 16, or a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 69, 71, or 74, and more preferably a gene consisting of the base sequence shown in SEQ ID NO: 1, 7, or 16, or a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 69, 71, or 74.

[0041] [cpr gene] The polypeptide encoded by the cpr gene is not particularly limited, and may have the function of donating electrons to the polypeptide encoded by the cyp gene to elicit the hydroxylation function of the polypeptide encoded by the cyp gene.

[0042] The origin of the cpr gene is not particularly limited, and examples thereof include the genera Aspergillus, Ustilago, Phanerochaete, Cochliobolus, Neurospora, and Rhodotorula. In one or more embodiments, the origin of the cpr gene is preferably the genera Aspergillus, Phanerochaete, Cochliobolus, Neurospora, and Rhodotorula, and more preferably the genera Aspergillus and Rhodotorula, from the viewpoint of improving resorcin productivity. Specific examples of species in each genus are the same as those of the cyp gene.

[0043] Examples of the cpr gene derived from Aspergillus niger include a gene consisting of the base sequence shown in SEQ ID NO:22, and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:76.

[0044] Examples of the cpr gene derived from Ustilago maydis include a gene consisting of the base sequence shown in SEQ ID NO:25, and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:77.

[0045] Examples of the cpr gene derived from Phanerochaete chrysosporium (Phanerodontia chrysosporium) include a gene consisting of the base sequence shown in SEQ ID NO:28 and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:78.

[0046] Examples of the cpr gene derived from Cochliobolus lunatus include a gene consisting of the base sequence shown in SEQ ID NO:31 and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:79.

[0047] Examples of the cpr gene derived from Neurospora crassa include a gene consisting of the base sequence shown in SEQ ID NO:34 and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:80.

[0048] Examples of the cpr gene derived from Rhodotorula minuta include a gene consisting of the nucleotide sequence shown in SEQ ID NO:37 and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:81.

[0049] The cpr gene is not particularly limited and may be, for example, any of the following genes (A7), (A8), and (A9). (A7) A gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 76, 77, 78, 79, 80 or 81, or a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 76, 77, 78, 79, 80 or 81. (A8) A gene encoding a polypeptide having an amino acid sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 76, 77, 78, 79, 80, or 81, and the gene encoding a polypeptide having a function of donating electrons to a polypeptide encoded by a cyp gene. or a gene encoding a polypeptide consisting of an amino acid sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 76, 77, 78, 79, 80 or 81, and which encodes a polypeptide having the function of donating electrons to a polypeptide encoded by the cyp gene. (A9) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 76, 77, 78, 79, 80 or 81, and which encodes a polypeptide having the function of donating electrons to a polypeptide encoded by the cyp gene, or a gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 76, 77, 78, 79, 80 or 81, and which encodes a polypeptide having the function of donating electrons to a polypeptide encoded by the cyp gene.

[0050] The cpr gene is not particularly limited and may be, for example, any of the following genes (A10), (A11) and (A12). (A10) A gene having a nucleotide sequence shown in SEQ ID NO: 22, 25, 28, 31, 34 or 37, or a gene consisting of a nucleotide sequence shown in SEQ ID NO: 22, 25, 28, 31, 34 or 37. (A11) A gene having a nucleotide sequence that has an identity of 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more to a gene having a nucleotide sequence shown in SEQ ID NO: 22, 25, 28, 31, 34, or 37, and which encodes a polypeptide having a function of donating electrons to a polypeptide encoded by a cyp gene. or a gene having a base sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the base sequence shown in SEQ ID NO: 22, 25, 28, 31, 34 or 37, and encoding a polypeptide having the function of donating electrons to a polypeptide encoded by the cyp gene. (A12) A gene that hybridizes under stringent conditions with a gene having a base sequence complementary to a gene having a base sequence shown in SEQ ID NO: 22, 25, 28, 31, 34 or 37, and encodes a polypeptide having the function of donating electrons to a polypeptide encoded by the cyp gene, or a gene that hybridizes under stringent conditions with a gene having a base sequence complementary to a gene consisting of the base sequence shown in SEQ ID NO: 22, 25, 28, 31, 34 or 37, and encodes a polypeptide having the function of donating electrons to a polypeptide encoded by the cyp gene.

[0051] In one or more embodiments, the cpr gene is preferably a cpr gene derived from Aspergillus niger from the viewpoint of improving resorcin productivity. From the same viewpoint, in one or more embodiments, the cpr gene is preferably a gene having the base sequence shown in SEQ ID NO: 22 or a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 76, and more preferably a gene consisting of the base sequence shown in SEQ ID NO: 22 or a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 76.

[0052] In one or more embodiments, the combination of the cyp gene and the cpr gene may be a combination of at least one of the cyp gene derived from Aspergillus niger, the cyp gene derived from Phanerochaete chrysosporium, and the cyp gene derived from Rhodotorula minuta with the cpr gene derived from Aspergillus niger, from the viewpoint of improving resorcinol productivity. In the same respect, the combination of the cyp gene and the cpr gene may be a combination of the cyp gene derived from Aspergillus niger with the cpr gene derived from Aspergillus niger, a combination of the cyp gene derived from Phanerochaete chrysosporium with the cpr gene derived from Aspergillus niger, or a combination of the cyp gene derived from Rhodotorula minuta with the cpr gene derived from Aspergillus niger. In a similar vein, combinations of the cyp gene and the cpr gene, in one or more embodiments, include a combination of a gene having a base sequence shown in SEQ ID NO: 1, 7 or 16 with a gene having a base sequence shown in SEQ ID NO: 22, or a combination of a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 69, 71 or 74 with a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 76. From a similar viewpoint, in one or more embodiments, the combination of the cyp gene and the cpr gene may be a combination of a gene having a nucleotide sequence shown in SEQ ID NO: 1 and a gene having a nucleotide sequence shown in SEQ ID NO: 22, a combination of a gene having a nucleotide sequence shown in SEQ ID NO: 7 and a gene having a nucleotide sequence shown in SEQ ID NO: 22, or a combination of a gene having a nucleotide sequence shown in SEQ ID NO: 16 and a gene having a nucleotide sequence shown in SEQ ID NO: 22. From a similar viewpoint, in one or more embodiments, the combination of the cyp gene and the cpr gene may be a combination of a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 69 and a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 76, a combination of a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 71 and a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 76, or a combination of a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 74 and a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 76.

[0053] [Gene (B)] In the present disclosure, the gene (B) may be one type of gene or a combination of two or more types of genes in one or more embodiments. In one or more embodiments, the gene (B) encoding an enzyme having 2,4-dihydroxybenzoic acid decarboxylation activity is not particularly limited, and examples thereof include genes encoding enzymes having decarboxylation activity for 2,4-dihydroxybenzoic acid, and genes encoding enzymes having activity to decarboxylate 2,4-dihydroxybenzoic acid to produce resorcinol are preferred. In one or more embodiments, examples of the gene (B) include decarboxylase genes ptaG, graF, sdc, and ubiD.

[0054] In one or more embodiments, whether or not the gene is a "gene encoding an enzyme having 2,4-dihydroxybenzoic acid decarboxylation activity" can be evaluated by carrying out a decarboxylation reaction of 2,4-dihydroxybenzoic acid using a transformant prepared by expressibly introducing the gene to be evaluated into Corynebacterium glutamicum. When the production or increase in the production amount of resorcin is confirmed by decarboxylating 2,4-dihydroxybenzoic acid, the gene can be determined to be a gene encoding an enzyme having 2,4-dihydroxybenzoic acid decarboxylation activity. The production of resorcin can be measured based on the description of the Examples in the present specification. In the present disclosure, the "gene encoding an enzyme having 2,4-dihydroxybenzoic acid decarboxylation activity" can also be said to be a gene encoding an enzyme having an activity of producing resorcin by decarboxylating 2,4-dihydroxybenzoic acid.

[0055] In one or more embodiments, from the viewpoint of improving resorcinol productivity, it is preferable that the enzyme encoded by gene (B) does not substantially have an activity of decarboxylating salicylic acid to produce phenol (also referred to as decarboxylase activity for salicylic acid). From the same viewpoint, it is preferable that the enzyme encoded by gene (B) has a decarboxylase activity for salicylic acid of 40% or less or 30% or less of the decarboxylase activity for 2,4-dihydroxybenzoic acid in one or more embodiments. The decarboxylase activity for salicylic acid can be evaluated by carrying out a decarboxylase reaction using salicylic acid as a substrate, using a transformant prepared by expressing a gene to be evaluated into Corynebacterium glutamicum. When the production of phenol is not substantially confirmed upon decarboxylation of salicylic acid, it can be determined that the sample does not substantially have salicylic acid decarboxylase activity. The production of phenol can be measured based on the description in the Examples of the present specification. In one or a plurality of embodiments, the decarboxylase activity for salicylic acid and the decarboxylase activity for 2,4-dihydroxybenzoic acid can be compared by carrying out the decarboxylase reaction using a transformant prepared by introducing a gene to be evaluated into Corynebacterium glutamicum so that the gene can be expressed, under the same culture conditions except for the substrate (salicylic acid or 2,4-dihydroxybenzoic acid), and comparing the products produced.

[0056] [ptaG gene] The origin of the ptaG gene is not particularly limited, and examples thereof include the genus Aspergillus. Examples of the genus Aspergillus are as described above, and among them, Aspergillus clavatus is preferred. Examples of the ptaG gene derived from Aspergillus clavatus include a gene consisting of the nucleotide sequence shown in SEQ ID NO: 43 and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 83.

[0057] The ptaG gene is not particularly limited and may be, for example, any one of the following genes (b1), (b2), and (b3). (b1) A gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 83, or a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 83. (b2) A gene encoding a polypeptide having an amino acid sequence that has 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 83, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity; or a gene encoding a polypeptide having an amino acid sequence that has 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 83, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity. (b3) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 83 and that encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity, or a gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 83 and that encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity.

[0058] The ptaG gene is not particularly limited and may be, for example, any one of the following genes (b4), (b5), and (b6). (b4) A gene having the base sequence shown in SEQ ID NO: 43, or a gene consisting of the base sequence shown in SEQ ID NO: 43. (b5) a gene having a nucleotide sequence that has an identity of 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more to the nucleotide sequence shown in SEQ ID NO: 43, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity; or a gene having a nucleotide sequence that has an identity of 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more to the nucleotide sequence shown in SEQ ID NO: 43, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity; (b6) A gene that hybridizes under stringent conditions with a gene having a nucleotide sequence complementary to the gene having the nucleotide sequence shown in SEQ ID NO: 43 and encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity, or a gene that hybridizes under stringent conditions with a gene having a nucleotide sequence complementary to the gene consisting of the nucleotide sequence shown in SEQ ID NO: 43 and encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity.

[0059] [graF gene] The origin of the graF gene is not particularly limited, and examples thereof include the genus Tsukamurella and the genus Mycobacterium.

[0060] The genus Tsukamurella according to the present disclosure is not particularly limited, and examples thereof include Tsukamurella paurometabola, Tsukamurella carboxydivorans, Tsukamurella tyrosinosolvens, Tsukamurella spongiae, and Tsukamurella poriferae, and may be Tsukamurella paurometabola. Examples of the graF gene derived from Tsukamurella paurometabola include a gene consisting of the base sequence shown in SEQ ID NO:46 and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:84.

[0061] The Mycobacterium genus according to the present disclosure is not particularly limited, for example, Mycobacterium immunogenum, Mycobacterium tuberculosis, Mycobacterium canettii, Mycobacterium leprae, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium paraintracellulare, Mycobacterium marseillense, Mycobacterium lepraemurium, Mycobacterium mantenii, Mycobacterium ulcerans, Mycobacterium marinum, Mycobacterium liflandii, Mycobacterium pseudoshottsii, Mycobacterium shottsii, Mycobacterium haemophilum, Mycobacterium kansasii, Mycobacterium dioxanotrophicus, Mycobacterium shigaense, Mycobacterium florentinum, Mycobacterium stomatepiae, Mycobacterium simiae, Mycobacterium saskatchewanense, Mycobacterium kubicae, Mycobacterium lentiflavum, Mycobacterium grossiae, Mycobacterium xenopi, Mycobacterium noviomagense, Mycobacterium paragordonae, Mycobacterium novum, Mycobacterium gordonae, Mycobacterium cookii, Mycobacterium basiliense, Mycobacterium seoulense, Mycobacterium heckeshornense, Mycobacterium gallinarum, Mycobacterium lacus, Mycobacterium branderi, MycobacteriumExamples of the Mycobacterium bacterium include Mycobacterium shinjukuense, Mycobacterium malmoense, Mycobacterium spongiae, Mycobacterium ostraviense, Mycobacterium lepromatosis, and Mycobacterium paraterrae, and may be Mycobacterium immunogenum. Examples of the graF gene derived from Mycobacterium immunogenum include a gene consisting of the nucleotide sequence shown in SEQ ID NO:49 and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:85.

[0062] The graF gene is not particularly limited and may be, for example, any of the following genes (b7), (b8), and (b9). (b7) A gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 84 or 85, or a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 84 or 85. (b8) a gene encoding a polypeptide having an amino acid sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 84 or 85, wherein the gene encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity. or a gene encoding a polypeptide consisting of an amino acid sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 84 or 85, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity. (b9) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 84 or 85, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity, or a gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 84 or 85, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity.

[0063] The graF gene is not particularly limited and may be, for example, any one of the following genes (b10), (b11), and (b12). (b10) A gene having the base sequence shown in SEQ ID NO: 46 or 49, or a gene consisting of the base sequence shown in SEQ ID NO: 46 or 49. (b11) a gene having a nucleotide sequence that has an identity of 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more to a gene having a nucleotide sequence represented by SEQ ID NO: 46 or 49, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity. or a gene having a nucleotide sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the nucleotide sequence shown in SEQ ID NO: 46 or 49, and encoding a polypeptide having 2,4-dihydroxybenzoic acid decarboxylase activity. (b12) A gene that hybridizes under stringent conditions with a gene having a nucleotide sequence complementary to a gene having the nucleotide sequence shown in SEQ ID NO: 46 or 49 and encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity, or a gene that hybridizes under stringent conditions with a gene having a nucleotide sequence complementary to a gene consisting of the nucleotide sequence shown in SEQ ID NO: 46 or 49 and encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity.

[0064] [sdc gene] The origin of the sdc gene is not particularly limited, and examples thereof include the genus Cutaneotrichosporon. The genus Cutaneotrichosporon according to the present disclosure is not particularly limited, and examples thereof include Cutaneotrichosporon moniliiforme, Cutaneotrichosporon oleaginosum, Cutaneotrichosporon dermatis, Cutaneotrichosporon curvatum, Cutaneotrichosporon mucoides, Cutaneotrichosporon cutaneum, and Cutaneotrichosporon jirovecii, and may be Cutaneotrichosporon moniliiforme. Examples of the sdc gene derived from Cutaneotrichosporon moniliiforme include a gene consisting of the base sequence shown in SEQ ID NO:52, and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:86.

[0065] The sdc gene is not particularly limited and may be, for example, any of the following genes (b13), (b14), and (b15). (b13) A gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 86, or a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 86. (b14) A gene encoding a polypeptide having an amino acid sequence that has 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 86, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity; or a gene encoding a polypeptide having an amino acid sequence that has 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 86, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity. (b15) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 86, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity, or a gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 86, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity.

[0066] The sdc gene is not particularly limited and may be, for example, any one of the following genes (b16), (b17), and (b18). (b16) A gene having the base sequence shown in SEQ ID NO: 52, or a gene consisting of the base sequence shown in SEQ ID NO: 52. (b17) A gene having a nucleotide sequence that has an identity of 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more to the nucleotide sequence shown in SEQ ID NO: 52, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity; or a gene having a nucleotide sequence that has an identity of 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more to the nucleotide sequence shown in SEQ ID NO: 52, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity. (b18) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene having the nucleotide sequence shown in SEQ ID NO: 52 and encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity, or a gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene consisting of the nucleotide sequence shown in SEQ ID NO: 52 and encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity.

[0067] [ubiD gene] The ubiD gene is a gene encoding a prenylated flavinine nucleotide (prFMN)-dependent decarboxylase, and the coenzyme prFMN is synthesized by a flavin prenyltransferase encoded by the ubiX gene. Thus, the polypeptide encoded by the ubiD gene, preferably in combination with the polypeptide encoded by the ubiX gene, can have 2,4-dihydroxybenzoic acid decarboxylation activity. The ubiD and ubiX genes may be under the control of a single promoter. It is known that the ubiH gene may exist in the same genome as the ubiD gene. From the viewpoint of improving resorcinol productivity, in one or more embodiments, it is preferable to introduce the ubiX gene in the same genome as the ubiD gene into the microbial host together with the ubiD gene. When the ubiH gene is present in the same genome as the ubiD gene, it is preferable to introduce the ubiH gene together with the ubiD gene, and it is more preferable to introduce the ubiH gene together with the ubiD gene and the ubiX gene, from the viewpoint of improving resorcinol productivity. Therefore, in one or more embodiments, examples of the gene (B) include the ubiDX gene, the ubiXD gene, the ubiDH gene, the ubiHD gene, the ubiXDH gene, the ubiHDX gene, the ubiDXH gene, the ubiDHX gene, the ubiXHD gene, and the ubiHXD gene, and the like, and from the viewpoint of improving resorcinol productivity, the ubiXDH gene, the ubiHDX gene, the ubiDXH gene, the ubiDHX gene, the ubiXHD gene, and the ubiHXD gene are preferable, and the ubiXDH gene is more preferable. In one or more embodiments, the ubiD gene and the ubiX gene and / or the ubiH gene may be introduced in combination with the ubiD gene and the ubiX gene and / or the ubiH gene of the same genus (or species), and it is not necessary to introduce in combination with the ubiD gene and the ubiX gene and / or the ubiH gene of the same genus (or species). Even when the ubiD gene and the ubiX gene and / or the ubiH gene of any different genera (or species) are introduced in combination into the host, it goes without saying that the ubiD gene has the activity of decarboxylating 2,4-dihydroxybenzoic acid to produce resorcinol. In one or more embodiments, the ubiD gene and the ubiX gene and / or the ubiH gene may be under the control of one promoter, or may be under the control of separate promoters. In the present disclosure, the term "ubiXHD genes" means that the ubiX gene, the ubiH gene, and the ubiD gene form an operon in this order.

[0068] The origin of the ubiD gene is not particularly limited, and examples thereof include the genus Pantoea and the genus Escherichia.

[0069] The genus Pantoea according to the present disclosure is not particularly limited, and examples thereof include Pantoea ananatis, Pantoea vagans, Pantoea rwandensis, Pantoea agglomerans, Pantoea stewartii, Pantoea alhagi, Pantoea eucalypti, Pantoea dispersa, Pantoea eucrina, Pantoea jilinensis, and Pantoea anthophila, and may be Pantoea ananatis. Examples of the ubiD gene derived from Pantoea ananatis include a gene consisting of the nucleotide sequence shown in SEQ ID NO:90, and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:87.

[0070] The genus Escherichia according to the present disclosure is not particularly limited, and examples thereof include Escherichia fergusonii, Escherichia coli, Escherichia albertii, Escherichia marmotae, and Escherichia ruysiae, and may be Escherichia fergusonii. Examples of the ubiD gene derived from Escherichia fergusonii include a gene consisting of the nucleotide sequence shown in SEQ ID NO:91, and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:88.

[0071] The ubiD gene is not particularly limited and may be, for example, any of the following genes (b19), (b20), and (b21). (b19) A gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 87 or 88, or a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 87 or 88. (b20) a gene encoding a polypeptide having an amino acid sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 87 or 88, wherein the gene encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity. or a gene encoding a polypeptide having an amino acid sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 87 or 88, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity. (b21) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 87 or 88, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity, or a gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 87 or 88, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity.

[0072] The ubiD gene is not particularly limited and may be, for example, any of the following genes (b22), (b23), and (b24). (b22) A gene having the nucleotide sequence shown in SEQ ID NO: 90 or 91, or a gene consisting of the nucleotide sequence shown in SEQ ID NO: 90 or 91. (b23) a gene having a nucleotide sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to a gene having a nucleotide sequence shown in SEQ ID NO: 90 or 91, and encoding a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity. or a gene having a nucleotide sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the nucleotide sequence shown in SEQ ID NO: 90 or 91, and encoding a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity; (b24) A gene that hybridizes under stringent conditions with a gene having a nucleotide sequence complementary to a gene having the nucleotide sequence shown in SEQ ID NO: 90 or 91 and encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity, or a gene that hybridizes under stringent conditions with a gene having a nucleotide sequence complementary to a gene consisting of the nucleotide sequence shown in SEQ ID NO: 90 or 91 and encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity.

[0073] [ubiX gene] The origin of the ubiX gene is not particularly limited, and examples thereof include the genus Pantoea and the genus Escherichia. Specific examples of species in the genus Pantoea and the genus Escherichia are the same as those of the ubiD gene. Examples of the ubiX gene derived from Pantoea ananatis include a gene consisting of the nucleotide sequence shown in SEQ ID NO:93, and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:98. Examples of the ubiX gene derived from Escherichia fergusonii include a gene consisting of the nucleotide sequence shown in SEQ ID NO:94, and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:99. The ubiX gene may form an operon with the ubiD gene and / or the ubiH gene, or may not form an operon.

[0074] The ubiX gene is not particularly limited and may be, for example, any of the following genes (b25), (b26), and (b27). (b25) A gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 98 or 99, or a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 98 or 99. (b26) a gene encoding a polypeptide having an amino acid sequence that has 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 98 or 99, and a gene encoding a polypeptide having flavin prenyltransferase activity, or a gene encoding a polypeptide consisting of an amino acid sequence that has 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 98 or 99, and a gene encoding a polypeptide having flavin prenyltransferase activity. (b27) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 98 or 99, and which encodes a polypeptide having flavin prenyltransferase activity, or a gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 98 or 99, and which encodes a polypeptide having flavin prenyltransferase activity. In one or more embodiments, the polypeptides encoded by the genes (b26) and (b27) may be capable of exhibiting 2,4-dihydroxybenzoic acid decarboxylation activity in combination with the polypeptide encoded by the ubiD gene.

[0075] The ubiX gene is not particularly limited and may be, for example, any of the following genes (b28), (b29) and (b30). (b28) a gene having the nucleotide sequence shown in SEQ ID NO: 93 or 94, or a gene consisting of the nucleotide sequence shown in SEQ ID NO: 93 or 94. (b29) a gene having a nucleotide sequence that has 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to a gene having a nucleotide sequence shown in SEQ ID NO: 93 or 94, and which encodes a polypeptide having flavin prenyltransferase activity; or a gene having a nucleotide sequence that has 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the nucleotide sequence shown in SEQ ID NO: 93 or 94, and which encodes a polypeptide having flavin prenyltransferase activity. (b30) A gene that hybridizes under stringent conditions with a gene having a nucleotide sequence complementary to a gene having the nucleotide sequence shown in SEQ ID NO: 93 or 94 and encodes a polypeptide having flavin prenyltransferase activity, or a gene that hybridizes under stringent conditions with a gene having a nucleotide sequence complementary to a gene consisting of the nucleotide sequence shown in SEQ ID NO: 93 or 94 and encodes a polypeptide having flavin prenyltransferase activity. In one or more embodiments, the polypeptides encoded by the genes (b29) and (b30) may be capable of exhibiting 2,4-dihydroxybenzoic acid decarboxylation activity in combination with a polypeptide encoded by the ubiD gene.

[0076] [ubiH gene] The origin of the ubiH gene is not particularly limited, and examples thereof include the genus Pantoea and the genus Escherichia. Specific examples of species in the genus Pantoea and the genus Escherichia are the same as those of the ubiD gene. Examples of the ubiH gene derived from Pantoea ananatis include a gene consisting of the nucleotide sequence shown in SEQ ID NO:96 and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:101. Examples of the ubiH gene derived from Escherichia fergusonii include a gene consisting of the nucleotide sequence shown in SEQ ID NO:97, and a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:102. The ubiH gene may form an operon with the ubiD gene and / or the ubiX gene, or may not form an operon.

[0077] The ubiH gene is not particularly limited and may be, for example, any of the following genes (b31), (b32), and (b33). (b31) A gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 101 or 102, or a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 101 or 102. (b32) a gene encoding a polypeptide having an amino acid sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 101 or 102, wherein the gene encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity in combination with a polypeptide encoded by the ubiD gene. or a gene encoding a polypeptide consisting of an amino acid sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 101 or 102, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity in combination with the polypeptide encoded by the ubiD gene. (b33) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 101 or 102, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity in combination with the polypeptide encoded by the ubiD gene, or a gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 101 or 102, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity in combination with the polypeptide encoded by the ubiD gene.

[0078] The ubiH gene is not particularly limited and may be, for example, any of the genes (b34), (b35), and (b36) below. (b34) A gene having the nucleotide sequence shown in SEQ ID NO: 96 or 97, or a gene consisting of the nucleotide sequence shown in SEQ ID NO: 96 or 97. (b35) a gene having a nucleotide sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to a gene having a nucleotide sequence shown in SEQ ID NO: 96 or 97, and encoding a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity in combination with a polypeptide encoded by the ubiD gene. or a gene having a nucleotide sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the nucleotide sequence shown in SEQ ID NO: 96 or 97, and encoding a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity in combination with a polypeptide encoded by the ubiD gene. (b36) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene having a nucleotide sequence shown in SEQ ID NO: 96 or 97, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity in combination with a polypeptide encoded by the ubiD gene, or a gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene consisting of the nucleotide sequence shown in SEQ ID NO: 96 or 97, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity in combination with a polypeptide encoded by the ubiD gene.

[0079] [ubiXDH gene] The origin of the ubiXDH gene is not particularly limited, and examples thereof include the genus Pantoea and the genus Escherichia. Specific examples of species in each genus are the same as those of the ubiD gene. An example of the ubiXDH gene derived from Pantoea ananatis is a gene consisting of the nucleotide sequence shown in SEQ ID NO: 55. The gene consisting of the nucleotide sequence shown in SEQ ID NO: 55 encodes a polypeptide (ubiX gene) consisting of the amino acid sequence shown in SEQ ID NO: 98, a polypeptide (ubiD gene) consisting of the amino acid sequence shown in SEQ ID NO: 87, and a polypeptide (ubiH gene) consisting of the amino acid sequence shown in SEQ ID NO: 101. An example of the ubiXDH gene derived from Escherichia fergusonii is a gene consisting of the nucleotide sequence shown in SEQ ID NO: 58. The gene consisting of the nucleotide sequence shown in SEQ ID NO: 58 encodes a polypeptide (ubiX gene) consisting of the amino acid sequence shown in SEQ ID NO: 99, a polypeptide (ubiD gene) consisting of the amino acid sequence shown in SEQ ID NO: 88, and a polypeptide (ubiH gene) consisting of the amino acid sequence shown in SEQ ID NO: 102.

[0080] The ubiXDH gene is not particularly limited and may be, for example, any of the following genes (b37), (b38) and (b39). (b37) A gene having the nucleotide sequence shown in SEQ ID NO: 55 or 58, or a gene consisting of the nucleotide sequence shown in SEQ ID NO: 55 or 58. (b38) a gene having a nucleotide sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to a gene having a nucleotide sequence represented by SEQ ID NO: 55 or 58, and encoding a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity. or a gene having a nucleotide sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the nucleotide sequence shown in SEQ ID NO: 55 or 58, and encoding a polypeptide having 2,4-dihydroxybenzoic acid decarboxylase activity. (b39) A gene that hybridizes under stringent conditions with a gene having a nucleotide sequence complementary to a gene having the nucleotide sequence shown in SEQ ID NO: 55 or 58 and encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity, or a gene that hybridizes under stringent conditions with a gene having a nucleotide sequence complementary to a gene consisting of the nucleotide sequence shown in SEQ ID NO: 55 or 58 and encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity.

[0081] In one or more embodiments, the gene (B) is not particularly limited and may be, for example, any of the following genes (B1), (B2), and (B3). (B1) A gene encoding a polypeptide having an amino acid sequence represented by SEQ ID NO: 83, 84, 85, 86, 87, or 88, or a gene encoding a polypeptide consisting of the amino acid sequence represented by SEQ ID NO: 83, 84, 85, 86, 87, or 88. (B2) A gene encoding a polypeptide having an amino acid sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 83, 84, 85, 86, 87, or 88, wherein the gene encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity. or a gene encoding a polypeptide consisting of an amino acid sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the amino acid sequence shown in SEQ ID NO: 83, 84, 85, 86, 87 or 88, and which encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity. (B3) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 83, 84, 85, 86, 87 or 88, and that encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity, or a gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 83, 84, 85, 86, 87 or 88, and that encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity.

[0082] In one or more embodiments, the gene (B) is not particularly limited and may be, for example, any of the following genes (B4), (B5), and (B6). (B4) A gene having a nucleotide sequence shown in SEQ ID NO: 43, 46, 49, 52, 90 or 91, or a gene consisting of a nucleotide sequence shown in SEQ ID NO: 43, 46, 49, 52, 90 or 91. (B5) A gene having a nucleotide sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to a gene having a nucleotide sequence represented by SEQ ID NO: 43, 46, 49, 52, 90, or 91, and encoding a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity. or a gene having a nucleotide sequence having 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.9% or more identity to the nucleotide sequence shown in SEQ ID NO: 43, 46, 49, 52, 90 or 91, and encoding a polypeptide having 2,4-dihydroxybenzoic acid decarboxylase activity. (B6) A gene that hybridizes under stringent conditions with a gene having a nucleotide sequence complementary to a gene having the nucleotide sequence shown in SEQ ID NO: 43, 46, 49, 52, 90 or 91 and encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity, or a gene that hybridizes under stringent conditions with a gene having a nucleotide sequence complementary to a gene consisting of the nucleotide sequence shown in SEQ ID NO: 43, 46, 49, 52, 90 or 91 and encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity.

[0083] As gene (B), in one or more embodiments, from the viewpoint of improving resorcinol productivity, the ptaG gene, the graF gene, and the ubiXDH gene are preferred, and from the same viewpoint, the ptaG gene derived from Aspergillus clavatus, the graF gene derived from Tsukamurella paurometabola, the ubiXDH gene derived from Pantoea ananatis, and the ubiXDH gene derived from Escherichia fergusonii are more preferred, and the ptaG gene derived from Aspergillus clavatus and the graF gene derived from Tsukamurella paurometabola are even more preferred.

[0084] In one or more embodiments, the combination of gene (A) and gene (B) may, from the viewpoint of improving resorcinol productivity, be a combination of at least one of the cyp gene derived from Aspergillus niger, the cyp gene derived from Phanerochaete chrysosporium, and the cyp gene derived from Rhodotorula minuta, and at least one of the cpr gene derived from Aspergillus niger, the ptaG gene derived from Aspergillus clavatus, the graF gene derived from Tsukamurella paurometabola, the ubiXDH gene derived from Pantoea ananatis, and the ubiXDH gene derived from Escherichia fergusonii, preferably a combination of at least one of the cyp gene derived from Aspergillus niger and the cyp gene derived from Rhodotorula minuta, the cpr gene derived from Aspergillus niger, and at least one of the ptaG gene derived from Aspergillus clavatus and the graF gene derived from Tsukamurella paurometabola. From a similar perspective, in one or more embodiments, combinations of gene (A) and gene (B) include a combination of a gene having the nucleotide sequence shown in SEQ ID NO: 1, 7 or 16, a gene having the nucleotide sequence shown in SEQ ID NO: 22, and a gene having the nucleotide sequence shown in SEQ ID NO: 43, 46, 55 or 58, and preferably a combination of a gene having the nucleotide sequence shown in SEQ ID NO: 1 or 16, a gene having the nucleotide sequence shown in SEQ ID NO: 22, and a gene having the nucleotide sequence shown in SEQ ID NO: 43 or 46. From a similar viewpoint, in one or more embodiments, the combination of gene (A) and gene (B) includes a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 69, 71 or 74, a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 76, a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 83, a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 84, a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 98, a polypeptide having an amino acid sequence shown in SEQ ID NO: 87, and a polypeptide having an amino acid sequence shown in SEQ ID NO: 101, or a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 99, a polypeptide having an amino acid sequence shown in SEQ ID NO: 88, and a polypeptide having an amino acid sequence shown in SEQ ID NO: 102, and preferably includes a combination of a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 69 or 74, a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 76, and a gene encoding a polypeptide having an amino acid sequence shown in SEQ ID NO: 83 or 84.

[0085] When gene (A) contains a cyp gene derived from Aspergillus niger, in one or more embodiments, gene (B) preferably contains at least one of the ptaG gene derived from Aspergillus clavatus, the graF gene derived from Tsukamurella paurometabola, the ubiXDH gene derived from Pantoea ananatis, and the ubiXDH gene derived from Escherichia fergusonii, from the viewpoint of improving resorcinol productivity, and more preferably contains at least one of the ptaG gene derived from Aspergillus clavatus and the graF gene derived from Tsukamurella paurometabola. From a similar standpoint, when gene (A) has a gene having the base sequence shown in SEQ ID NO: 1, in one or more embodiments, gene (B) preferably includes at least one of the genes having the base sequences shown in SEQ ID NOs: 43, 46, 55 and 58, and more preferably includes at least one of the genes having the base sequences shown in SEQ ID NOs: 43 and 46, in terms of improving resorcinol productivity. From a similar standpoint, when gene (A) has a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO:69, in one or more embodiments, from the standpoint of improving resorcinol productivity, gene (B) preferably includes at least one of a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO:83, a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO:84, a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO:98, a polypeptide having the amino acid sequence shown in SEQ ID NO:87, and a polypeptide having the amino acid sequence shown in SEQ ID NO:101, and a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO:99, a polypeptide having the amino acid sequence shown in SEQ ID NO:88, and a polypeptide having the amino acid sequence shown in SEQ ID NO:102, and more preferably includes at least one of genes encoding polypeptides having the amino acid sequences shown in SEQ ID NOs:83 and 84.

[0086] When the gene (A) contains the cyp gene derived from Rhodotorula minuta, in one or more embodiments, the gene (B) preferably contains at least one of the ptaG gene derived from Aspergillus clavatus and the graF gene derived from Tsukamurella paurometabola, from the viewpoint of improving resorcinol productivity. From a similar standpoint, when gene (A) has a gene having the base sequence shown in SEQ ID NO: 16, in one or more embodiments, gene (B) preferably includes a gene having the base sequence shown in SEQ ID NO: 43 or 46 in terms of improving resorcinol productivity. From a similar standpoint, when gene (A) has a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 74, in one or more embodiments, gene (B) preferably includes a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 83 or 84 in terms of improving resorcinol productivity.

[0087] In one embodiment, from the viewpoint of improving resorcinol productivity, the transformant according to the present disclosure may have the cyp gene, cpr gene, ptaG gene, graF gene, sdc gene or ubiXDH gene introduced in an expressible manner into a microbial host capable of producing salicylic acid, preferably the cyp gene, cpr gene, ptaG gene, graF gene or ubiXDH gene are introduced in an expressible manner, and more preferably the cyp gene, cpr gene, ptaG gene or graF gene are introduced in an expressible manner.

[0088] In one aspect, the transformant according to the present disclosure may have enhanced resorcinol production from sugar by increasing the production of salicylic acid, which is an endogenous precursor as a reaction substrate for the introduced gene (A).

[0089] Although not essential, in one embodiment, the transformant according to the present disclosure may optionally have enhanced activity of one or more enzymes contained in at least one of the shikimic acid pathway, the sugar metabolic pathway (iolT1-ppgk), and the nonoxidative pentose phosphate pathway. The enhancement of the enzyme activity is not particularly limited, and may be achieved, for example, by enhancing the expression of the gene encoding the enzyme.

[0090] Salicylic acid is a biosynthetic intermediate of tryptophan, an aromatic amino acid, and is produced by a catalytic reaction of salicylic acid synthase using chorismic acid, which is produced in the shikimic acid pathway, a common metabolic pathway involved in aromatic amino acid production, as a substrate. For this reason, the metabolic flux of the shikimic acid pathway may be increased in order to increase the production of resorcin. FIG. 2 is a schematic diagram of an example of an entire sugar metabolic pathway showing the metabolic pathway of resorcin using sugars as a raw material. The sugar metabolic pathway in the present disclosure refers to various in vivo reaction systems for decomposing and converting sugars such as glucose to raw materials for the biosynthesis of various compounds, and in one or more embodiments, may include the shikimic acid pathway, the sugar metabolic pathway (iolT1-ppgk), the nonoxidative pentose phosphate pathway, and the TCA cycle. However, the biosynthetic pathway of resorcin shown in FIG. 2 is merely an example of the sugar metabolic pathway in the present disclosure, and the present disclosure is not limited thereto.

[0091] The metabolic engineering technique for increasing the metabolic flux of the shikimic acid pathway is not particularly limited, and in one embodiment, it may be any of the following pathways (1) to (3), or a combination of two or three of pathways (1) to (3). (1) High activation of the enzyme genes that compose the shikimate pathway (aroG, aroB, aroD, aroE, aroK, aroA, and aroC, which code for 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase, 3-dehydroquinate synthase, 3-dehydroquinate dehydratase, shikimate dehydrogenase, shikimate kinase, 5-enolpyruvylshikimate-3-phosphate synthase, and chorismate synthase, respectively) by enhancing their expression or using heterologous or mutant genes. (2) High activation by enhancing the expression of the genes constituting the non-PTS sugar transport pathway, which is an alternative sugar transport and metabolic pathway different from the phosphoenolpyruvate:sugar phosphotransferase system (PTS) commonly used as a sugar transporter in wild-type strains (iolT1 and iolT2 genes encoding myo-inositol / glucose transporters, glk gene encoding glucokinase, and ppgk gene encoding polyphosphate glucokinase). (3) High activation of the genes encoding the enzymes that make up the nonoxidative pentose phosphate pathway (tkt and tal, encoding transketolase and transaldolase, respectively) by enhancing their expression. In one or more embodiments, the metabolic modification may be, but is not limited to, at least one modification of a gene constituting each pathway, or may be two, three, or four or more modifications of a gene constituting each pathway, or may be modifications of all of the genes mentioned above in each pathway.

[0092] In order to improve resorcin productivity, the transformant according to the present disclosure may have at least one or all of the aroG gene, aroD gene, aroE gene, aroC gene, aroK gene, and aroB gene introduced into the host.

[0093] Although not essential, in one embodiment, the transformant according to the present disclosure may optionally have reduced or deleted functions of, for example, a gene encoding 3-dehydroshikimate dehydratase (e.g., qsuB gene), quinic acid / shikimate dehydrogenase (e.g., qsuD gene), and lactate dehydrogenase (e.g., ldhA gene). From the viewpoint of improving resorcinol productivity, the transformant according to the present disclosure may have a part or whole of the qsuB gene, qsuD gene, and ldhA gene in the host disrupted or deleted.

[0094] In one or more embodiments, the transformant according to the present disclosure may have lost, inhibited, or reduced activity of an enzyme that uses resorcin as a substrate, from the viewpoint of improving resorcin productivity. In one or more embodiments, the transformant according to the present disclosure may have a part or all of a gene encoding an enzyme that uses resorcin as a substrate disrupted or deleted, from the viewpoint of improving resorcin productivity. In one or more embodiments, examples of genes encoding enzymes using resorcinol as a substrate include the mhpA gene (enzyme encoded: EC number 1.14.13.219, reosrcinol 4-hydroxylase (NADPH)), the tsdB gene (enzyme encoded: EC number 1.14.13.220, reosrcinol 4-hydroxylase (NADH)), the graA gene (enzyme encoded: EC number 1.14.14.27, reosrcinol 4-hydroxylase (FADH2)), and the phe gene (enzyme encoded: EC number 1.14.13.7, phenol 2-monooxygenase (NADPH)). In one or more embodiments, the transformant according to the present disclosure may have lost, inhibited, or reduced activity of the enzymes using resorcinol as a substrate, which are encoded by the mhpA gene, the tsdB gene, the graA gene, and the phe gene, from the viewpoint of improving resorcinol productivity.

[0095] In one embodiment, the transformant according to the present disclosure may be modified so that each metabolic pathway is strengthened by enhancing the expression of the various metabolic pathway genes or by highly activating enzyme functions through the use of heterologous (mutated) genes.

[0096] [Vector / plasmid] In another aspect, the present disclosure relates to a vector for introducing the above gene (A) into a microorganism.In another aspect, the present disclosure relates to a vector for introducing the above gene (B) into a microorganism. In yet another aspect, the present disclosure relates to a vector for introducing a cyp gene and a cpr gene into a microorganism.In yet another aspect, the present disclosure relates to a vector for introducing at least one of a ptaG gene, a graF gene, an sdc gene, and an ubiXDH gene into a microorganism, preferably a vector for introducing at least one of a ptaG gene, a graF gene, and an sdc gene into a microorganism, more preferably a vector for introducing at least one of a ptaG gene and a graF gene into a microorganism. The vector according to the present disclosure is not particularly limited and may be, for example, a plasmid.

[0097] Construction of vectors for transformation The above genes can be introduced into a microbial host by amplifying the above-described genes by PCR, cloning them into an appropriate vector that can be amplified in a microbial host such as a coryneform bacterium, and incubating the microbial host in the presence of the vector. In one or more embodiments, examples of the promoter include the promoter (PgapA) of the gapA gene encoding glyceraldehyde 3-phosphate dehydrogenase (also referred to as "glyceraldehyde 3-phosphate dehydrogenase") derived from Corynebacterium glutamicum R, the promoter (Pmdh) of the mdh gene encoding malate dehydrogenase, and the promoter (PldhA) of the ldhA gene encoding lactate dehydrogenase, with PgapA being preferred.

[0098] [Creation of transformants] Transformation The transformation method can be any known method without limitation. In one or more embodiments, known methods include the calcium chloride / rubidium chloride method, the calcium phosphate method, DEAE-dextran mediated transfection, and electroporation (electric pulse method). When the microbial host is a coryneform bacterium, the electric pulse method is preferable. The electric pulse method can be performed by known methods [e.g., Kurusu, Y. et al., Electroporation-transformation system for Coryneform bacteria by auxotrophic complementation. Agric. Biol. Chem. 54:443-447(1990)] and [Vertes AA et al., Presence of mrr- and mcr-like restriction systems in Coryneform bacteria. Res. Microbiol. 144:181-185(1993)].

[0099] Disruption or mutation of host chromosomal genes When the microbial host is a coryneform bacterium, genes encoding competing pathways for biosynthesis, inhibitors of the biosynthetic pathway, or efflux transporters may be disrupted and / or deleted as necessary. The function of an enzyme protein encoded by a specific gene may be improved by introducing a mutation into a chromosome. A DNA fragment in which the entire length of a target gene is deleted is prepared by linking DNA fragments before and after the target gene, and a bacterium is transformed with the DNA to cause homologous recombination on the chromosome, thereby completely deleting the target gene on the chromosome. Alternatively, a deletion-type gene modified so that a partial sequence of the target gene is deleted and a normally functioning enzyme protein is not produced is prepared, and a bacterium is transformed with DNA containing the gene to cause homologous recombination between the deletion-type gene and the gene on the chromosome, thereby replacing the target gene on the chromosome with a deletion-type or disruption-type gene. Even if an enzyme protein encoded by a deletion-type or disruption-type gene is produced, it has a three-dimensional structure different from that of a wild-type enzyme protein, and its function is reduced or lost. In addition, a mutation can be introduced into a specific position on a chromosome by causing homologous recombination between a gene fragment in which a specific mutation has been introduced and the chromosomal region. Such gene deletion or disruption by gene replacement using homologous recombination has already been established, and includes methods using plasmids containing a temperature-sensitive replication origin, plasmids capable of conjugative transfer, and methods using suicide vectors that do not have a replication origin in the host (U.S. Pat. No. 6,303,383, JP 05-007491 A). The markerless chromosomal gene introduction vector pCRA725 is a plasmid that cannot replicate in Corynebacterium glutamicum R. In the case of a single-crossover strain with a homologous region on the chromosome introduced into the plasmid pCRA725, the strain exhibits kanamycin resistance due to the expression of the kanamycin resistance gene on pCRA725 and lethality in a sucrose-containing medium due to the expression of the sacR-sacB gene of Bacillus subtilis, whereas in the case of a double-crossover strain, the strain exhibits kanamycin sensitivity due to the loss of the kanamycin resistance gene on pCRA725 and growth in a sucrose-containing medium due to the loss of the sacR-sacB gene. Thus, the markerless chromosomal gene introduction strain exhibits kanamycin sensitivity and growth in a sucrose-containing medium.

[0100] Microbial growth In one or more embodiments, the transformant according to the present disclosure is preferably cultured and grown under aerobic conditions prior to reaction in a reaction solution. In one or more embodiments, the culture conditions are a temperature of about 25° C. to 38° C. and a time of about 12 hours to 48 hours. In one or more embodiments, the medium used for the aerobic culture may be a natural medium or a synthetic medium containing a carbon source, a nitrogen source, inorganic salts, other nutrients, etc. In one or more embodiments, the pH of the medium is preferably about 5 to 8.

[0101] Examples of media for coryneform bacteria include A medium [Inui, M. et al., Metabolic analysis of Corynebacterium glutamicum during lactate and succinate productions under oxygen deprivation conditions. J. Mol. Microbiol. Biotechnol. 7:182-196(2004)] and BT medium [Omumasaba, CA et al., Corynebacterium glutamicum glyceraldehyde-3-phosphate dehydrogenase isoforms with opposite, ATP-dependent regulation. J. Mol. Microbiol. Biotechnol. 8:91-103(2004)]. In one or more embodiments, the medium for E. coli may be LB medium or the like.

[0102] In one or more embodiments, the carbon source may be a carbohydrate or sugar alcohol such as glucose, fructose, sucrose, mannose, maltose, mannitol, xylose, arabinose, galactose, starch, molasses, sorbitol, or glycerin; an organic acid such as acetic acid, citric acid, lactic acid, fumaric acid, maleic acid, or gluconic acid; or an alcohol such as ethanol or propanol. If desired, a hydrocarbon such as normal paraffin may also be used. The carbon source may be used alone or in combination of two or more. In one or more embodiments, the concentration of the carbon source in the growth medium is about 0.1 (w / v%) to 10 (w / v%).

[0103] In one or more embodiments, examples of the nitrogen source include inorganic or organic ammonium compounds such as ammonium chloride, ammonium sulfate, ammonium nitrate, and ammonium acetate, urea, aqueous ammonia, sodium nitrate, and potassium nitrate. In one or more embodiments, examples of the nitrogen source include nitrogen-containing organic compounds such as corn steep liquor, meat extract, peptone, NZ-amine, protein hydrolysates, and amino acids. The nitrogen source may be used alone or in combination of two or more. The concentration of the nitrogen source in the growth medium varies depending on the nitrogen compound used, but is about 0.1 (w / v%) to 10 (w / v%) in one or more embodiments.

[0104] In one or more embodiments, examples of inorganic salts include monopotassium phosphate, dipotassium phosphate, magnesium sulfate, sodium chloride, ferrous nitrate, manganese sulfate, zinc sulfate, cobalt sulfate, ammonium carbonate, calcium carbonate, etc. One type of inorganic salt may be used alone, or two or more types may be mixed and used. The concentration of inorganic salts in the growth medium varies depending on the inorganic salt used, but in one or more embodiments, it is about 0.01 (w / v%) to 1 (w / v%).

[0105] In one or more embodiments, examples of other nutritional substances include meat extract, peptone, polypeptone, yeast extract, dry yeast, corn steep liquor, skimmed milk powder, hydrolyzed skimmed soybean with hydrochloric acid, or extracts of animals, plants, or microbial cells, or decomposition products thereof. The concentration of the nutritional substances in the medium varies depending on the nutritional substances used, but in one or more embodiments, it is about 0.1 (w / v%) to 10 (w / v%). Vitamins may be added as necessary. In one or more embodiments, examples of vitamins include biotin, thiamine (vitamin B1), pyridoxine (vitamin B6), pantothenic acid, inositol, and nicotinic acid.

[0106] [Method of producing resorcinol] The present disclosure relates to a method for producing resorcinol. In detail, in one aspect, the method for producing resorcinol according to the present disclosure may be a production method including a step of culturing a transformant according to the present disclosure in a reaction solution containing sugars to produce resorcinol.

[0107] The reaction liquid may contain saccharides. In one or more embodiments, the reaction liquid may be water, a buffer solution, an inorganic salt medium, the above-mentioned medium, or the like, containing saccharides. In one or more embodiments, the reaction liquid may contain nutrients other than saccharides, such as vitamins, yeast extract, and dry yeast.

[0108] In one or more embodiments, the sugars include glucose, fructose, mannose, xylose, arabinose, galactose, sucrose, maltose, lactose, cellobiose, xylobiose, trehalose, mannitol, etc. In one or more embodiments, the concentration of the sugars in the reaction solution is about 0.1 (w / v%) to 20 (w / v%), and preferably 1 (w / v%) to 20 (w / v%) or 5 (w / v%) to 20 (w / v%).

[0109] In one or more embodiments, the buffer solution may be a phosphate buffer, a tris buffer, a carbonate buffer, etc. In one or more embodiments, the concentration of the buffer solution may be about 10 mM to 150 mM.

[0110] In one or more embodiments, the inorganic salt medium may include a medium containing one or more inorganic salts such as potassium monophosphate, potassium diphosphate, molybdenum sulfate, sodium chloride, ferrous nitrate, manganese sulfate, zinc sulfate, cobalt sulfate, calcium carbonate, urea, ammonium sulfate, and ferrous sulfate. Among them, a medium containing urea, ammonium sulfate, potassium monophosphate, potassium diphosphate, magnesium sulfate, and ferrous sulfate is preferable. Specific examples of the inorganic salt medium include BT medium and A medium. The concentration of inorganic salts in the inorganic salt medium varies depending on the inorganic salt used, but in one or more embodiments, the concentration is about 0.01 (w / v%) to 1 (w / v%).

[0111] Reaction conditions In one or more embodiments, the reaction conditions include reducing conditions and microaerobic conditions (for example, conditions in which the dissolved oxygen concentration is controlled), etc. In one or more embodiments, which are not particularly limited, when the microbial host is Corynebacterium glutamicum, the reaction is carried out under both reducing conditions and microaerobic conditions in which Corynebacterium glutamicum does not substantially grow, so that resorcin can be produced more efficiently.

[0112] In one or more embodiments, the reaction temperature (the survival temperature of the transformant during the reaction) is about 15° C. to 50° C. Within the above temperature range, resorcin can be produced efficiently. From the viewpoint of improving resorcin productivity, the temperature is 16° C. or higher, 17° C. or higher, 18° C. or higher, 19° C. or higher, or 20° C. or higher. From the same viewpoint, the temperature is 49° C. or lower, 45° C. or lower, 40° C. or lower, 35° C. or lower, 34° C. or lower, or 33° C. or lower.

[0113] In one or more embodiments, the pH of the reaction solution is preferably about 6 to 8. During the reaction, it is preferable to control the pH of the reaction solution to near neutral, particularly about 7, using an aqueous ammonia solution, an aqueous sodium hydroxide solution, or the like with a pH controller (e.g., Model: DT-1023, manufactured by Able Co., Ltd.).

[0114] In one or more embodiments, the reaction time is about 1 to 7 days, and preferably about 1 to 3 days. In one or more embodiments, the culture may be any of a batch system, a fed-batch system, and a continuous system, and among these, the batch system is preferred.

[0115] In one or more embodiments, the method for producing resorcin of the present disclosure may include recovering resorcin from the reaction solution (culture), and may further include purifying the recovered resorcin as necessary. The method for recovering and purifying resorcin from the reaction solution (culture) is not particularly limited. In one or more embodiments, the recovery and purification can be carried out by appropriately combining well-known ion exchange resin methods, precipitation methods, crystallization methods, recrystallization methods, concentration methods, and other methods. Resorcin accumulated in the culture may be used as it is without isolation.

[0116] The present disclosure further relates to one or more of the following embodiments. [1] A transformant obtained by introducing the following genes (A) and (B) into a microbial host in an expressible manner: (A) A gene encoding an enzyme that hydroxylates salicylic acid to produce 2,4-dihydroxybenzoic acid. (B) A gene encoding an enzyme having 2,4-dihydroxybenzoic acid decarboxylation activity. [2] The transformant according to [1], wherein the gene (A) is derived from at least one selected from the group consisting of the genera Aspergillus, Ustilago, Phanerochaete, Cochliobolus, Neurospora, and Rhodotorula. [3] The transformant according to [1] or [2], wherein the gene (A) includes at least a gene selected from the group consisting of (A1), (A2) and (A3) below. (A1) a gene encoding a polypeptide having the amino acid sequence represented by SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, or SEQ ID NO: 74; (A2) a gene encoding a polypeptide having an amino acid sequence having 85% or more identity to the amino acid sequence shown in SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, or SEQ ID NO: 74, and having an activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid; (A3) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, or SEQ ID NO:74, and that encodes a polypeptide having the activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid. [4] The transformant described in [1] or [2], wherein the gene (A) includes at least a gene selected from the group consisting of (A4), (A5) and (A6) below. (A4) a gene having the nucleotide sequence represented by SEQ ID NO: 1, SEQ ID NO: 4, SEQ ID NO: 7, SEQ ID NO: 10, SEQ ID NO: 13, or SEQ ID NO: 16; (A5) a gene having a nucleotide sequence having 85% or more identity to the nucleotide sequence shown in SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:16, which encodes a polypeptide having an activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid; (A6) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene having the nucleotide sequence shown in SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13 or SEQ ID NO:16, and that encodes a polypeptide having the activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid. [5] The transformant described in any of [1] to [4], wherein the gene (B) is derived from at least one selected from the group consisting of the genera Aspergillus, Tsukamurella, Mycobacterium, Cutaneotrichosporon, Pantoea, and Escherichia. [6] The transformant according to any one of [1] to [5], wherein the gene (B) includes at least a gene selected from the group consisting of (B1), (B2) and (B3) below. (B1) a gene encoding a polypeptide having the amino acid sequence represented by SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, or SEQ ID NO: 88; (B2) a gene encoding a polypeptide having an amino acid sequence having 85% or more identity to the amino acid sequence shown in SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, or SEQ ID NO: 88, wherein the gene encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity; (B3) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, or SEQ ID NO: 88, and that encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity. [7] The transformant described in any of [1] to [5], wherein the gene (B) includes at least a gene selected from the group consisting of (B4), (B5) and (B6) below. (B4) a gene having the nucleotide sequence represented by SEQ ID NO: 43, SEQ ID NO: 46, SEQ ID NO: 49, SEQ ID NO: 52, SEQ ID NO: 90, or SEQ ID NO: 91; (B5) a gene having a nucleotide sequence having 85% or more identity to the nucleotide sequence shown in SEQ ID NO: 43, SEQ ID NO: 46, SEQ ID NO: 49, SEQ ID NO: 52, SEQ ID NO: 90, or SEQ ID NO: 91, and encoding a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity; (B6) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene having the nucleotide sequence shown in SEQ ID NO: 43, SEQ ID NO: 46, SEQ ID NO: 49, SEQ ID NO: 52, SEQ ID NO: 90, or SEQ ID NO: 91, and that encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity. [8] The transformant according to any of [1] to [7], wherein the gene (B) encodes a decarboxylase that has substantially no decarboxylase activity against salicylic acid, or encodes a decarboxylase that has higher decarboxylase activity against 2,4-dihydroxybenzoic acid than the decarboxylase activity against salicylic acid. [9] The transformant according to any one of [1] to [8], wherein the microbial host is a coryneform bacterium.

[10] The transformant according to any one of [1] to [9], wherein the microbial host is Corynebacterium glutamicum.

[11] The transformant according to any one of [1] to

[10] , wherein the microbial host is Corynebacterium glutamicum R (FERM BP-18976), ATCC13032, ATCC13869, or a transformant thereof.

[12] The transformant described in any one of [1] to

[11] , wherein the microbial host has the ability to produce salicylic acid.

[13] A microbial host capable of producing salicylic acid The cyp gene encoding cytochrome P450; The cpr gene, which encodes cytochrome P450 reductase; At least one selected from the group consisting of a ptaG gene encoding a decarboxylase, a graF gene, a sdc gene, and a ubiD gene, A transformant obtained by introducing an expression vector.

[14] the cyp gene and the cpr gene are derived from at least one selected from the group consisting of the genera Aspergillus, Cochliobolus, Phanerochaete, Neurospora, and Rhodotorula; and / or The transformant according to

[13] , wherein the gene encoding the decarboxylase is derived from at least one selected from the group consisting of the genera Aspergillus, Tsukamurella, Mycobacterium, Cutaneotrichosporon, Pantoea, and Escherichia.

[15] The cyp gene is at least one selected from the group consisting of (A1), (A2), (A3), (A4), (A5) and (A6) above, and / or The cpr gene is at least one selected from the group consisting of (A7), (A8), (A9), (A10), (A11) and (A12) above, and / or The transformant according to

[13] or

[14] , wherein the gene encoding a decarboxylase is at least one selected from the group consisting of (B1), (B2), (B3), (B4), (B5) and (B6).

[16] The transformant according to any one of

[13] to

[15] , wherein the microbial host is a coryneform bacterium.

[17] The transformant according to any one of

[13] to

[16] , wherein the microbial host is Corynebacterium glutamicum.

[18] The transformant according to any one of

[13] to

[17] , wherein the microbial host is Corynebacterium glutamicum R (FERM BP-18976), ATCC13032, ATCC13869, or a transformant thereof.

[19] The transformant described in any one of [1] to

[18] , which has the ability to produce resorcinol.

[20] A method for producing resorcinol, comprising a step of culturing the transformant described in any one of [1] to

[19] in the presence of a sugar to produce resorcinol.

[0117] The present disclosure will be further described below using examples, but the present disclosure is not limited to the following examples. EXAMPLES

[0118] [Example 1] Construction of resorcinol-producing strain (1) Preparation and acquisition of chromosomal DNA Chromosomal DNA of Amycolatopsis methanolica JCM 8087, Aspergillus clavatus JCM 1718, Aspergillus niger JCM 22282, Corynebacterium glutamicum R (FERM BP-18976), Cutaneotrichosporon moniliiforme NBRC 1527, Escherichia fergusonii NBRC 102419, Mycobacterium immunogenum JCM 12691, Neurospora crassa ATCC 36373, Pantoea ananatis LMG 20103, Trichoderma reesei NBRC 31329, and Tsukamurella paurometabola ATCC 8368 was extracted with a DNA genome extraction kit (illustra bacteria genomicPrep Mini Spin) after culturing according to the information provided by the institution where the strain was obtained. The chromosomal DNA of Rhodobacter capsulatus ATCC BAA-309 was obtained from ATCC. The hydroxylase genes of Cochliobolus lunatus, Phanerochaete chrysosporium, Rhodotorula minuta, and Ustilago maydis, and the decarboxylase gene of Rhodotorula minuta were prepared by gene synthesis.

[0119] (2) Construction of expression plasmid for resorcinol production-related genes The primer sequences used to isolate the target enzyme genes are shown in Table 1. PCR was performed using a VeritiPro thermal cycler (Thermo Fisher Scientific Co., Ltd.) and PrimeSTAR HS DNA Polymerase (Takara Bio Inc.) as a reaction reagent. [Table 1]

[0120] The PCR-amplified DNA fragment was introduced into the cloning vector pCRB209 [International Publication WO2012 / 033112] containing the PgapA promoter. The cpr gene was introduced as a PgapA promoter fusion gene fragment into the plasmid pCRB1 [NCBI GenBank: AB444682] that can coexist in Corynebacterium glutamicum, and the cyp gene and the cpr gene were introduced as a PgapA promoter fusion gene fragment into pCRB11. The obtained plasmids are shown in Table 2. [Table 2]

[0121] (3) Construction of a plasmid for introducing resorcinol production-related genes into chromosome The DNA region required for markerless introduction of a resorcinol production-related gene into the chromosome of Corynebacterium glutamicum R strain was determined based on a sequence reported to be non-essential for the growth of Corynebacterium glutamicum R strain [Appl. Environ. Microbiol. 71:3369-3372(2005)] (SSI region). This DNA region was amplified by PCR, and the resulting DNA fragment was introduced into the markerless gene introduction plasmid pCRA725 [J. Mol. Microbiol. Biotechnol. 8:243-254(2004), JP 2007-295809 A]. The primer sequences and the resulting plasmids are shown in Table 3. [Table 3] The PgapA promoter fusion gene fragment was obtained from the resorcinol production-related gene expression plasmid constructed in Table 2 and introduced into the above-mentioned plasmid for chromosomal introduction. The obtained plasmid is shown in Table 4. [Table 4]

[0122] (4) Construction of strains with resorcinol-producing genes introduced into the chromosome The markerless chromosomal gene introduction vector pCRA725 is a plasmid that cannot replicate in Corynebacterium glutamicum R. In the case of a single-crossover strain with a homologous region on the chromosome introduced into the plasmid pCRA725, the strain exhibits kanamycin resistance due to the expression of the kanamycin resistance gene on pCRA725 and lethality in a sucrose-containing medium due to the expression of the sacR-sacB gene derived from Bacillus subtilis, whereas in the case of a double-crossover strain, the strain exhibits kanamycin sensitivity due to the loss of the kanamycin resistance gene on pCRA725 and growth in a sucrose-containing medium due to the loss of the sacR-sacB gene. Thus, the markerless chromosomal gene introduction strain exhibits kanamycin sensitivity and growth in a sucrose-containing medium. By the above method, a genetically modified strain was constructed using the above-mentioned plasmid for chromosomal introduction of resorcinol production-related genes. The host strain used was the Corynebacterium glutamicum R ldhA-disrupted strain CRZ1 [Biotechnol Bioeng. Nov;110(11):2938-2948(2013)]. The plasmid pCRB284 [WO2017 / 169399] for aroG gene chromosome introduction, the plasmid pCRB291 [WO2017 / 169399] for aroD gene chromosome introduction, the plasmid pCRB293 [WO2017 / 169399] for aroE gene chromosome introduction, the plasmid pCRB286 [WO2017 / 169399] for aroCKB gene chromosome introduction, the plasmid pCRB229 [WO2017 / 169399] for qsuB gene disruption, and the plasmid pCRB299 [WO2017 / 169399] for qsuD gene disruption were also used. The overview of this chromosomal gene recombination is summarized in Table 5. [Table 5]

[0123] (5) Construction of a strain expressing a resorcinol-producing gene The above-mentioned resorcin production-related gene expression plasmid was introduced into Corynebacterium glutamicum R strain and Rrsol strain. The overview of the introduced strains is summarized in Table 6. [Table 6]

[0124] [Experiment (1)] Hydroxylase screening experiment (an experiment to hydroxylate salicylic acid to produce 2,4-dihydroxybenzoic acid (2,4-DHBA)) The hydroxylase gene-introduced strains (RSSK01-RSSK06) constructed in Example 1 were used to examine the ability of each strain to produce 2,4-dihydroxybenzoic acid from salicylic acid. Each strain was cultured in 10 ml of ACK-Glc liquid medium (A liquid medium (2 g (NH2)2CO, 7 g (NH4)2SO4, 0.5 g KH2PO4, 0.5 g K2HPO4, 0.5 g MgSO4·7H2O, 1 ml of a mixture of 0.6% (w / v) FeSO4·7H2O and 0.42% (w / v) MnSO4·H2O, 1 ml of 0.02% (w / v) biotin solution, 1 ml of 0.02% (w / v) thiamin solution, 2 g of yeast extract, and vitamin C assay casamino acid. The bacteria was pre-cultured overnight in a liquid medium (7 g dissolved in 1 L of distilled water) (in a test tube) containing 4% glucose, 5 μg / ml chloramphenicol, and 50 μg / ml kanamycin. The bacteria was then inoculated into 10 ml of fresh ACK-Glc liquid medium containing 10 mM salicylic acid to an initial OD of 0.2, and the reaction was carried out at 20°C for 48 hours with shaking. The detection area of ​​2,4-dihydroxybenzoic acid contained in the reaction supernatant of each strain was measured by HPLC analysis, and the amount of 2,4-dihydroxybenzoic acid produced by each strain using salicylic acid as a substrate was calculated as an activity measurement value. As a result, the amount of 2,4-dihydroxybenzoic acid produced by each strain is shown in Table 7. [Table 7]

[0125] The hydroxylase activity of the above strains against benzoic acid was measured. Each strain was cultured in 10 ml of ACK-Glc liquid medium (A liquid medium ((NH2)2CO 2g, (NH4)2SO4 7g, KH2PO4 0.5g, K2HPO4 0.5g, MgSO4·7H2O 0.5g, 0.6% (w / v) FeSO4·7H2O and 0.42% (w / v) MnSO4·H2O mixture 1ml, 0.02% (w / v) biotin solution 1ml, 0.02% (w / v) thiamin solution 1ml, yeast extract 2g, and vitamin assay casamino acid 1ml. The bacteria was pre-cultured overnight in a liquid medium (in a test tube) containing 7 g of the bacteria dissolved in 1 L of distilled water, 4% glucose, 5 μg / ml chloramphenicol, and 50 μg / ml kanamycin, and then inoculated into 10 ml of fresh ACK-Glc liquid medium containing 10 mM benzoic acid to an initial OD of 0.2, and the reaction was carried out at 20°C for 48 hours with shaking. The detection area of ​​4-hydroxybenzoic acid contained in the reaction supernatant of each strain was measured by HPLC analysis, and the amount of 4-hydroxybenzoic acid produced by each strain using benzoic acid as a substrate was calculated. As a result, the amount of 4-hydroxybenzoic acid produced by each strain was as follows. For the strain RSSK01 (derived from Aspergillus niger), it was 31.0 μM. For the strain RSSK02 (derived from Ustilago maydis), it was 5.2 μM. For the strain RSSK03 (derived from Phanerochaete chrysosporium), it was 16.8 μM. The strain RSSK04 (derived from Cochliobolus lunatus) had a concentration of 27.1 μM. For the strain RSSK05 (derived from Neurospora crassa), it was 5.4 μM. For the strain named RSSK06 (derived from Rhodotorula minuta), it was 8.5 μM. Therefore, it can be said that the hydroxylase encoded by gene (A) may have hydroxylase activity for benzoic acid. The hydroxylase encoded by gene (A) may have a hydroxylase activity for benzoic acid that is higher than the hydroxylase activity for salicylic acid, or may have a hydroxylase activity for salicylic acid that is lower than the hydroxylase activity for salicylic acid.

[0126] [Experiment (2)] Decarboxylase screening experiment (experiment to produce resorcinol by decarboxylating 2,4-DHBA) The decarboxylase gene-introduced strains (RDTS01-RDTS06) constructed in Example 1 were used to examine the ability of each strain to produce resorcinol from 2,4-DHBA. Each strain was pre-cultured overnight in 10 ml of AK-Glc liquid medium (liquid medium A (2 g of (NH2)2CO, 7 g of (NH4)2SO4, 0.5 g of KH2PO4, 0.5 g of K2HPO4, 0.5 g of MgSO4·7H2O, 1 ml of a mixture of 0.6% (w / v) FeSO4·7H2O and 0.42% (w / v) MnSO4·2H2O, 1 ml of 0.02% (w / v) biotin solution, 1 ml of 0.02% (w / v) thiamin solution, 2 g of yeast extract, and 7 g of vitamin assay casamino acid dissolved in 1 L of distilled water) supplemented with 4% glucose and 50 μg / ml kanamycin) (in a test tube). 0.5 ml of the culture was then taken and 0.5 ml of the reaction solution (20 mM The mixture was mixed with 2,4-DHBA, 200 mM 2-Morpholinoethanesulfonic acid, and the reaction was carried out by shaking at 33°C for 3 hours. The resorcinol contained in the reaction supernatant of each strain was analyzed by HPLC to measure the detection area and determine the resorcinol production concentration (measured activity). As a result, the resorcinol production concentration (measured activity) of each strain was as shown in Table 8. [Table 8]

[0127] The decarboxylase gene-introduced strains (RDTS01, RDT02, RDT03, RDT05, and RDT06) constructed in Example 1 were used to measure decarboxylase activity against salicylic acid. Each strain was pre-cultured overnight in 10 ml of AK-Glc liquid medium (liquid medium A (2 g (NH2)2CO, 7 g (NH4)2SO4, 0.5 g KH2PO4, 0.5 g K2HPO4, 0.5 g MgSO4·7H2O, 1 ml of a mixture of 0.6% (w / v) FeSO4·7H2O and 0.42% (w / v) MnSO4·2H2O, 1 ml of 0.02% (w / v) biotin solution, 1 ml of 0.02% (w / v) thiamin solution, 2 g of yeast extract, and 7 g of vitamin assay casamino acid dissolved in 1 L of distilled water) supplemented with 4% glucose and 50 μg / ml kanamycin) (in a test tube). 0.5 ml of the culture was then taken and 0.5 ml of the reaction solution (20 mM salicylic acid, 200 mM The mixture was mixed with 2-Morpholinoethanesulfonic acid and the reaction was carried out by shaking at 33°C for 3 hours. The detection area of ​​salicylic acid contained in the supernatant of the reaction solution of each strain was measured by HPLC analysis. As a result, the decarboxylase activity for salicylic acid was as follows. For the strain RDTS01 (derived from Aspergillus clavatus), 0.0 mM, For the strain named RDTS02 (derived from Tsukamurella paurometabola), it was 0.9 mM, For the strain named RDTS03 (derived from Mycobacterium immunogenum), it was 1.6 mM, For strain name RDTS05 (derived from Pantoea ananatis), 0.0 mM, For the strain named RDTS06 (derived from Escherichia fergusonii), it was 0.0 mM. Therefore, it is preferable that the decarboxylase encoded by gene (B) has substantially no decarboxylase activity for salicylic acid, or even if it has decarboxylase activity for salicylic acid, the activity is 40% or less or 30% or less compared to the decarboxylase activity for 2,4-DHBA.

[0128] [Experiment (3)] Resorcinol production from sugars Using the strains into which the hydroxylase gene and decarboxylase gene were introduced constructed in Example 1, the ability of each strain to produce resorcinol from sugar was examined. Each strain was produced in 10 ml of ACK-Glc liquid medium (liquid medium A (2 g of (NH2)2CO, 7 g of (NH4)2SO4, 0.5 g of KH2PO4, 0.5 g of K2HPO4, 0.5 g of MgSO4·7H2O, 1 ml of a mixture of 0.6% (w / v) FeSO4·7H2O and 0.42% (w / v) MnSO4·2H2O, 1 ml of 0.02% (w / v) biotin solution, 1 ml of 0.02% (w / v) thiamin solution, 2 g of yeast extract, and 7 g of vitamin assay casamino acid dissolved in 1 L of distilled water) supplemented with 4% glucose, 5 μg / ml of chloramphenicol, and 50 μg / ml of kanamycin) (in a test tube) with shaking at 20°C for 7 days. The resorcinol contained in the culture supernatant of each strain was measured by HPLC analysis to determine the detection area and the resorcinol production concentration. The results are shown in Table 9. As shown in Table 9, the resorcinol production concentrations of the strains RITK01, RITK02, RITK05, RITK06, RITK08, RITK11 and RITK12 were particularly high. [Table 9]

[0129] The abbreviated compound names and their CAS numbers shown in Figure 2 (an overall metabolic pathway diagram for explaining the biosynthetic pathway of resorcin) are shown below. It goes without saying that the "compound names" shown below are only examples, and there may be synonymous aliases. It goes without saying that the "CAS numbers" shown below are not necessarily a comprehensive list. [Table 10]

[0130] The names of the enzymes encoded by the genes shown in Figure 2 (an overall metabolic pathway diagram for explaining the biosynthetic pathway of resorcinol) and their EC numbers are shown below. The "enzyme names" shown below are merely examples, and it goes without saying that there may be synonymous alternative names. The "enzymes" encoded by the "genes" shown below are merely examples, and it goes without saying that there may be "genes" that encode "enzymes" with multiple functions. [Table 11]

Claims

1. A transformant obtained by introducing the following genes (A) and (B) into a microbial host in an expressible manner: (A) A gene encoding an enzyme having the activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid. (B) A gene encoding an enzyme having 2,4-dihydroxybenzoic acid decarboxylation activity.

2. The transformant according to claim 1, wherein the gene (A) is derived from at least one selected from the group consisting of the genera Aspergillus, Ustilago, Phanerochaete, Cochliobolus, Neurospora, and Rhodotorula.

3. The transformant according to claim 1 , wherein the gene (A) includes at least a gene selected from the group consisting of the following (A1), (A2) and (A3): (A1) a gene encoding a polypeptide having an amino acid sequence represented by SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, or SEQ ID NO: 74; (A2) a gene encoding a polypeptide having an amino acid sequence having 85% or more identity to the amino acid sequence shown in SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, or SEQ ID NO:74, and having an activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid; (A3) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, or SEQ ID NO:74, and that encodes a polypeptide having the activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid.

4. The transformant according to claim 1 , wherein the gene (A) includes at least a gene selected from the group consisting of the following genes (A4), (A5) and (A6): (A4) a gene having a nucleotide sequence represented by SEQ ID NO: 1, SEQ ID NO: 4, SEQ ID NO: 7, SEQ ID NO: 10, SEQ ID NO: 13, or SEQ ID NO: 16; (A5) a gene having a nucleotide sequence having 85% or more identity to the nucleotide sequence shown in SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:16, which encodes a polypeptide having an activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid; (A6) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene having the nucleotide sequence shown in SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:16, and that encodes a polypeptide having the activity of hydroxylating salicylic acid to produce 2,4-dihydroxybenzoic acid.

5. 2. The transformant according to claim 1, wherein the gene (B) is derived from at least one selected from the group consisting of the genera Aspergillus, Tsukamuralla, Mycobacterium, Cutaneotrichosporon, Pantoea, and Escherichia.

6. The transformant according to claim 1 , wherein the gene (B) includes at least a gene selected from the group consisting of the following (B1), (B2) and (B3): (B1) a gene encoding a polypeptide having an amino acid sequence represented by SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, or SEQ ID NO: 88; (B2) a gene encoding a polypeptide having an amino acid sequence having 85% or more identity to the amino acid sequence represented by SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, or SEQ ID NO: 88, and encoding a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity; (B3) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene encoding a polypeptide having the amino acid sequence shown in SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, or SEQ ID NO:88, and that encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity.

7. The transformant according to claim 1 , wherein the gene (B) includes at least a gene selected from the group consisting of the following genes (B4), (B5) and (B6): (B4) a gene having a nucleotide sequence represented by SEQ ID NO: 43, SEQ ID NO: 46, SEQ ID NO: 49, SEQ ID NO: 52, SEQ ID NO: 90, or SEQ ID NO: 91; (B5) a gene having a nucleotide sequence having 85% or more identity to the nucleotide sequence represented by SEQ ID NO: 43, SEQ ID NO: 46, SEQ ID NO: 49, SEQ ID NO: 52, SEQ ID NO: 90, or SEQ ID NO: 91, and encoding a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity; (B6) A gene that hybridizes under stringent conditions to a gene having a nucleotide sequence complementary to a gene having the nucleotide sequence shown in SEQ ID NO:43, SEQ ID NO:46, SEQ ID NO:49, SEQ ID NO:52, SEQ ID NO:90, or SEQ ID NO:91, and encodes a polypeptide having 2,4-dihydroxybenzoic acid decarboxylation activity.

8. The transformant according to claim 1, wherein the gene (B) encodes a decarboxylase having substantially no decarboxylase activity against salicylic acid, or encodes a decarboxylase having a decarboxylase activity against 2,4-dihydroxybenzoic acid higher than the decarboxylase activity against salicylic acid.

9. The transformant according to claim 1 , wherein the microbial host is a coryneform bacterium.

10. The transformant according to claim 1 , wherein the microbial host is Corynebacterium glutamicum.

11. The transformant according to claim 1, wherein the microbial host is Corynebacterium glutamicum R (FERM BP-18976), ATCC13032, ATCC13869, or a transformant thereof.

12. The transformant according to claim 1 , wherein the microbial host has an ability to produce salicylic acid.

13. A microbial host capable of producing salicylic acid The cyp gene encoding cytochrome P450; The cpr gene encoding cytochrome P450 reductase; At least one selected from the group consisting of a ptaG gene encoding a decarboxylase, a graF gene, a sdc gene, and a ubiD gene, A transformant obtained by introducing an expression vector.

14. The transformant according to claim 1 , which has resorcinol producing ability.

15. A method for producing resorcin, comprising a step of culturing the transformant according to any one of claims 1 to 14 in the presence of a saccharide to produce resorcin.

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