Novel lipase

Abstract

To provide a lipase whose activity is less likely to be inhibited by surfactants and which exhibits a high cleaning effect.SOLUTION: Provided is a lipase comprising an amino acid sequence represented by SEQ ID NO: 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42 or 44 or an amino acid sequence having at least substantial identity to any one of these amino acid sequences.SELECTED DRAWING: None

Description

[Technical Field] 【0001】 This invention relates to a novel lipase. [Background technology] 【0002】 Lipase is useful in a variety of applications, including laundry detergents, dish soaps, oil and fat processing, pulp processing, animal feed, and pharmaceutical intermediate synthesis. In cleaning, lipase contributes to the removal of oily dirt by hydrolyzing triglycerides to produce fatty acids. 【0003】 Current cleaning compositions and environments contain various components that inhibit lipase activity and cleaning effectiveness, and there is a need for lipases that function under such conditions. As a lipase useful for cleaning, lipase derived from Thermomyces lanuginosus (hereinafter referred to as TLL) is sold under the trade name LIPOLASE®. Patent Document 1 discloses that lipase Lipr139 derived from Cedecea sp-16640 strain has superior cleaning performance compared to TLL. Patent Document 2 discloses a variant of lipase derived from Proteus bacteria (hereinafter referred to as PvLip) that has improved cleaning performance compared to one or more reference lipolytic enzymes. Patent Document 3 discloses that metagenomic lipase Lipr138 has superior cleaning performance compared to TLL. Lipr139, PvLip, and Lipr138 are lipases belonging to the same clade (Proteus / Yersinia clade) which includes lipases derived from Proteus and Yersinia bacteria. 【0004】 Many lipases derived from Gram-negative bacteria require lipase-specific chaperones for folding into their active form (Non-Patent Literature 1), posing challenges to inexpensive production through heterologous expression. While co-expression of specific chaperones has been reported to improve heterologous expression (Non-Patent Literature 2), its productivity remains low, posing significant challenges to inexpensive production through heterologous expression. On the other hand, bacterial lipases from the Proteus / Yersinia clade do not require specific chaperones, offering advantages in inexpensive production through heterologous expression (Non-Patent Literature 3, 4). As described above, bacterial lipases from the Proteus / Yersinia clade have excellent potential as washing enzymes, both in terms of their suitability for washing and their low-cost production potential. [Prior art documents] [Patent Documents] 【0005】 [Patent Document 1] Special Publication No. 2015-523078 [Patent Document 2] International Publication No. 2020 / 046613 [Patent Document 3] Special Publication No. 2015-525248 [Non-patent literature] 【0006】 [Non-Patent Document 1] Hobson, Audrey H., et al. Proceedings of the National Academy of Sciences 90.12 (1993): 5682-5686. [Non-Patent Document 2] Quyen, ThiDinh, ChiHai Vu, and GiangThi Thu Le. Microbial cell factories 11.1 (2012): 1-12. [Non-Patent Document 3] Lee, Hong-Weon, et al. Biotechnology letters 22.19 (2000): 1543-1547. [Non-Patent Document 4] Glogauer, Arnaldo, et al. Microbial cell factories 10.1 (2011): 1-15. [Overview of the Initiative] [Problems that the invention aims to solve] 【0007】 It is known that the activity of lipases is inhibited by surfactants contained in cleaning compositions. In fact, as far as the present inventors have verified, all known lipases disclosed as suitable for cleaning—TLL, Lipr139, and PvLip—showed significant activity inhibition in the presence of surfactants. Activity inhibition by surfactants is a major challenge common to all cleaning lipases. Furthermore, lipases are often used with surfactants not only in cleaning applications but also in industrial applications such as pulp processing, so activity inhibition by surfactants is a challenge for industrial lipases as a whole. Therefore, there is a need for a lipase that exhibits high cleaning efficacy while mitigating activity inhibition by surfactants. [Means for solving the problem] 【0008】 In view of these problems, the inventors conducted diligent research and, as a result, discovered that a novel group of lipase sequences, presumed to have been present in the ancestral organisms of existing lipases belonging to the Proteus / Yersinia clade, surprisingly exhibits remarkably high resistance to activity inhibition by surfactants and possesses outstanding cleaning performance. While ancestral enzymes are generally known to exhibit properties such as high heat resistance and high optimal temperatures, their properties regarding inhibition of activity by surfactants and their cleaning action at low temperatures were completely unknown until now, making the above results unexpected. 【0009】 In other words, the present invention relates to the following 1) to 6). 1) Any of the following lipases: A lipase comprising the amino acid sequence shown in Sequence ID No. 14 or an amino acid sequence having at least 79% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 16 or an amino acid sequence having at least 76% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 18 or an amino acid sequence having at least 81% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 20 or an amino acid sequence having at least 83% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 22 or an amino acid sequence having at least 96% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 24 or an amino acid sequence having at least 83% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 26 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 28 or an amino acid sequence having at least 82% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 30 or an amino acid sequence having at least 73% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 32 or an amino acid sequence having at least 73% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 34 or an amino acid sequence having at least 79% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 36 or an amino acid sequence having at least 85% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 38 or an amino acid sequence having at least 73% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 40 or an amino acid sequence having at least 71% identity thereto; A lipase consisting of the amino acid sequence represented by SEQ ID NO: 42 or an amino acid sequence having at least 72% identity thereto; and A lipase consisting of the amino acid sequence represented by SEQ ID NO: 44 or an amino acid sequence having at least 70% identity thereto. 2) A polynucleotide encoding the lipase according to 1). 3) A vector or DNA fragment containing the polynucleotide according to 2). 4) A transformed cell containing the vector or DNA fragment according to 3). 5) A detergent composition containing the lipase according to 1). 6) A method for cleaning dirt, using the detergent composition according to 5). 【Advantages of the Invention】 【0010】 The lipase of the present invention has significantly high resistance to activity inhibition by surfactants and exhibits excellent cleaning effects even in the presence of surfactants. 【Brief Description of the Drawings】 【0011】 [Figure 1] Lipase activity of each lipase in a surfactant solution. [Figure 2] Lipase activity of each lipase in a model cleaning solution containing a surfactant. [Figure 3] Cleaning power of each lipase in a model cleaning solution containing a surfactant. [Figure 4] Results of phylogenetic analysis of each lipase. 【Modes for Carrying Out the Invention】 【0012】 All patent documents, non-patent documents, and other publications cited in this specification are hereby incorporated by reference in their entirety. 【0013】 In this specification, "lipase" refers to triacylglycerol lipase (EC 3.1.1.3), and means a group of enzymes that have the activity to hydrolyze triglycerides to produce fatty acids. Lipase activity can be determined by measuring the rate of increase in absorbance associated with the release of 4-nitrophenol by hydrolysis of 4-nitrophenyl octanoate. The specific procedure for measuring lipase activity is described in detail in the examples below. 【0014】 In this specification, "lipase presumed to have been possessed by the ancestral organism" means a lipase (also simply called ancestral lipase) consisting of an amino acid sequence presumed to have been possessed by the common ancestor, derived from the amino acid sequences of lipases of each organism derived from the common ancestor, based on a rooted phylogenetic tree representing the evolution of proteins. Ancestor lipases can be designed by obtaining lipase homolog sequences from public databases, performing multiple alignments using the obtained homolog sequences, then creating a rooted phylogenetic tree, and estimating the amino acid sequence of the common ancestor lipase using an ancestral sequence reconstruction (ASR) program. 【0015】 General methods can be used to design ancestral lipases, such as those described in Merkl R, Sterner R., Biol Chem. 2016;397(1):1-21 and Scossa F, Fernie AR., Comput Struct Biotechnol J. 2021;19:1579-1594. General phylogenetic analysis methods described in JSBi Bioinformatics Review, 2(1), 30-57 (2021) can also be used. Lipase homolog sequence datasets can be obtained from public databases such as NCBI and UniProtKB using BLAST (Altschul et al., 1990). Alternatively, they can be obtained in bulk as families from domain databases such as InterPro and Pfam. Multiple alignments can be created using programs such as Clustal X and Clustal W (Larkin et al., 2007), MUSCLE (Edgar, 2004), MAFFT (Katoh and Standley, 2013), MAFFT-DASH (Rozewicki et al., 2019), PRANK (Loytynoja, 2014), and T-COFFEE (Notredame et al., 2000). The created multiple alignments are then appropriately trimmed using programs such as GBLOCKS (Castresana, 2000) or trimAl (Capella-Gutierrez et al., 2009) before being used to estimate phylogenetic trees. For phylogenetic tree estimation, parsimony, Bayesian methods, and maximum likelihood methods can be used. For example, programs such as MrBayes (Ronquist et al., 2012), BEAST (Bouckaert et al., 2019), FastTree (Price et al., 2010), PhyML (Guindon et al., 2010), RAxML (Stamatakis et al., 2014), and IQ-TREE (Nguyen et al., 2015) can be used.Appropriate evolutionary models for phylogenetic tree estimation can be selected using programs such as modeltest-ng (Darriba, D. et al., 2020) and ModelFinder (Kalyaanamoorthy et al., 2017). Dedicated programs can be used for ancestral sequence design, including FastML (Pupko et al., 2000), ProtASR2 (Arenas M and Bastolla, 2020), and GRASP (Gabriel et al., 2019). Some phylogenetic tree estimation programs also include ancestral sequence design as a function, and programs such as PAML (Yang, 1997), RAxML, and IQ-TREE can also be used. MEGA (Tamura et al., 2021) can be used as an integrated environment to perform ancestral sequence design from these phylogenetic analyses in one go. 【0016】 In this specification, the "Proteus / Yersinia clade" refers to a clade on a rooted phylogenetic tree that includes lipases derived from bacteria of the genus Proteus and Yersinia (e.g., SEQ ID NOs: 4 and 12), and the amino acid sequences shown in SEQ ID NOs: 2, 4, 6, 8, 10, 12 and 46 are all extant lipase sequences belonging to the Proteus / Yersinia clade. 【0017】 In this specification, the identity of amino acid sequences or nucleotide sequences is calculated using the Lipman-Pearson method (Science, 1985, 227:1435-1441). Specifically, it is calculated by performing the homology analysis (Search homology) using the genetic information processing software GENETYX Ver.12 with a Unit size to compare (ktup) of 2. 【0018】 In this specification, "operable linkage" between a regulatory region such as a promoter and a gene means that the gene and the regulatory region are linked in such a way that the gene can be expressed under the control of the regulatory region. Procedures for "operable linkage" between a gene and a regulatory region are well known to those skilled in the art. 【0019】 In this specification, "upstream" and "downstream" with respect to a gene refer to the upstream and downstream regions in the transcription direction of the gene. For example, "a gene located downstream of a promoter" means that the gene is located on the 3' side of the promoter in the DNA sense strand, and "upstream of a gene" means the 5' region of the gene in the DNA sense strand. 【0020】 <1. Lipase> The lipase of the present invention is any of the following lipases: A lipase comprising the amino acid sequence shown in Sequence ID No. 14 or an amino acid sequence having at least 79% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 16 or an amino acid sequence having at least 76% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 18 or an amino acid sequence having at least 81% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 20 or an amino acid sequence having at least 83% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 22 or an amino acid sequence having at least 96% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 24 or an amino acid sequence having at least 83% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 26 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 28 or an amino acid sequence having at least 82% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 30 or an amino acid sequence having at least 73% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 32 or an amino acid sequence having at least 73% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 34 or an amino acid sequence having at least 79% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 36 or an amino acid sequence having at least 85% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 38 or an amino acid sequence having at least 73% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 40 or an amino acid sequence having at least 71% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 42 or an amino acid sequence having at least 72% identity thereto; and A lipase comprising the amino acid sequence shown in Sequence ID No. 44 or an amino acid sequence having at least 70% identity thereto, That is the case. Here, the amino acid sequences represented by sequence numbers 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, and 44 are novel lipase sequences that are presumed to have been present in the ancestral organisms of the bacteria from which existing lipases belonging to the Proteus / Yersinia clade originated. Generally, ancestral enzymes are known to exhibit properties such as high heat resistance and high optimal temperatures, but their properties regarding inhibition of activity by surfactants and their cleaning action have been completely unknown until now. It was completely unexpected that the lipase of the present invention has remarkably high resistance to inhibition of lipase activity by surfactants and exhibits remarkably high cleaning power even in the presence of surfactants. 【0021】 Lipases comprising an amino acid sequence having at least 79% identity with the amino acid sequence shown in SEQ ID NO: 14 include lipases comprising an amino acid sequence having at least 79% identity with the amino acid sequence shown in SEQ ID NO: 14, specifically, 79% or more, preferably 82% or more, more preferably 85% or more, even more preferably 90% or more, even more preferably 95% or more, even more preferably 96% or more, even more preferably 97% or more, even more preferably 98% or more, and even more preferably 99% or more identity. Amino acid sequences having at least 79% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0022】 Lipases comprising an amino acid sequence having at least 76% identity with the amino acid sequence shown in SEQ ID NO: 16 include lipases comprising an amino acid sequence having at least 76% identity with the amino acid sequence shown in SEQ ID NO: 16, specifically, 76% or more, preferably 80% or more, more preferably 85% or more, even more preferably 90% or more, even more preferably 95% or more, even more preferably 96% or more, even more preferably 97% or more, even more preferably 98% or more, and even more preferably 99% or more. Amino acid sequences having at least 76% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0023】 Lipases comprising an amino acid sequence having at least 81% identity with the amino acid sequence shown in SEQ ID NO: 18 include lipases comprising an amino acid sequence having at least 81% identity with the amino acid sequence shown in SEQ ID NO: 18, specifically, 81% or more, preferably 85% or more, more preferably 90% or more, even more preferably 95% or more, even more preferably 96% or more, even more preferably 97% or more, even more preferably 98% or more, and even more preferably 99% or more. Amino acid sequences having at least 81% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0024】 Lipases comprising an amino acid sequence having at least 83% identity with the amino acid sequence shown in SEQ ID NO: 20 include lipases comprising an amino acid sequence having at least 83% identity with the amino acid sequence shown in SEQ ID NO: 20, specifically, 83% or more, preferably 85% or more, more preferably 90% or more, even more preferably 95% or more, even more preferably 96% or more, even more preferably 97% or more, even more preferably 98% or more, and even more preferably 99% or more. Amino acid sequences having at least 83% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0025】 Lipases comprising an amino acid sequence having at least 96% identity with the amino acid sequence shown in SEQ ID NO: 22 include lipases comprising an amino acid sequence having at least 96% identity with the amino acid sequence shown in SEQ ID NO: 22, specifically, 96% or more, preferably 97% or more, more preferably 98% or more, and even more preferably 99% or more identity. Amino acid sequences having at least 96% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0026】 Lipases comprising an amino acid sequence having at least 83% identity with the amino acid sequence shown in SEQ ID NO: 24 include lipases comprising an amino acid sequence having at least 83% identity with the amino acid sequence shown in SEQ ID NO: 24, specifically, 83% or more, preferably 85% or more, more preferably 90% or more, even more preferably 95% or more, even more preferably 96% or more, even more preferably 97% or more, even more preferably 98% or more, and even more preferably 99% or more identity. Amino acid sequences having at least 83% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0027】 Lipases comprising an amino acid sequence having at least 90% identity with the amino acid sequence shown in SEQ ID NO: 26 include lipases comprising an amino acid sequence having at least 90% identity with the amino acid sequence shown in SEQ ID NO: 26, specifically, 90% or more, preferably 93% or more, more preferably 95% or more, even more preferably 96% or more, even more preferably 97% or more, even more preferably 98% or more, and even more preferably 99% or more. Amino acid sequences having at least 90% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0028】 Lipases comprising an amino acid sequence having at least 82% identity with the amino acid sequence shown in SEQ ID NO: 28 include lipases comprising an amino acid sequence having at least 83% identity with the amino acid sequence shown in SEQ ID NO: 28, specifically, 82% or more, preferably 85% or more, more preferably 90% or more, even more preferably 95% or more, even more preferably 96% or more, even more preferably 97% or more, even more preferably 98% or more, and even more preferably 99% or more. Amino acid sequences having at least 82% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0029】 Lipases comprising an amino acid sequence having at least 73% identity with the amino acid sequence shown in SEQ ID NO: 30 include lipases comprising an amino acid sequence having at least 73% identity with the amino acid sequence shown in SEQ ID NO: 30, specifically, 73% or more, preferably 75% or more, more preferably 80% or more, even more preferably 85% or more, even more preferably 90% or more, even more preferably 95% or more, even more preferably 96% or more, even more preferably 97% or more, even more preferably 98% or more, and even more preferably 99% or more identity. Amino acid sequences having at least 73% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0030】 Lipases comprising an amino acid sequence having at least 73% identity with the amino acid sequence shown in SEQ ID NO: 32 include lipases comprising an amino acid sequence having at least 73% identity with the amino acid sequence shown in SEQ ID NO: 32, specifically, 73% or more, preferably 75% or more, more preferably 80% or more, even more preferably 85% or more, even more preferably 90% or more, even more preferably 95% or more, even more preferably 96% or more, even more preferably 97% or more, even more preferably 98% or more, and even more preferably 99% or more. Amino acid sequences having at least 73% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0031】 Lipases comprising an amino acid sequence having at least 79% identity with the amino acid sequence shown in SEQ ID NO: 34 include lipases comprising an amino acid sequence having at least 79% identity with the amino acid sequence shown in SEQ ID NO: 34, specifically, 79% or more, preferably 82% or more, more preferably 85% or more, even more preferably 90% or more, even more preferably 95% or more, even more preferably 96% or more, even more preferably 97% or more, even more preferably 98% or more, and even more preferably 99% or more identity. Amino acid sequences having at least 79% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0032】 Lipases comprising an amino acid sequence having at least 85% identity with the amino acid sequence shown in SEQ ID NO: 36 include lipases comprising an amino acid sequence having at least 85% identity with the amino acid sequence shown in SEQ ID NO: 36, specifically, 85% or more, preferably 90% or more, more preferably 95% or more, even more preferably 96% or more, even more preferably 97% or more, even more preferably 98% or more, and even more preferably 99% or more. Amino acid sequences having at least 85% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0033】 Lipases comprising an amino acid sequence having at least 73% identity with the amino acid sequence shown in SEQ ID NO: 38 include lipases comprising an amino acid sequence having at least 73% identity with the amino acid sequence shown in SEQ ID NO: 38, specifically, 73% or more, preferably 75% or more, more preferably 80% or more, even more preferably 85% or more, even more preferably 90% or more, even more preferably 95% or more, even more preferably 96% or more, even more preferably 97% or more, even more preferably 98% or more, and even more preferably 99% or more. Amino acid sequences having at least 73% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0034】 Lipases comprising an amino acid sequence having at least 71% identity with the amino acid sequence shown in SEQ ID NO: 40 include lipases comprising an amino acid sequence having at least 71% identity with the amino acid sequence shown in SEQ ID NO: 40, specifically, 71% or more, preferably 75% or more, more preferably 80% or more, even more preferably 85% or more, even more preferably 90% or more, even more preferably 95% or more, even more preferably 96% or more, even more preferably 97% or more, even more preferably 98% or more, and even more preferably 99% or more. Amino acid sequences having at least 71% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0035】 Lipases comprising an amino acid sequence having at least 72% identity with the amino acid sequence shown in SEQ ID NO: 42 include lipases comprising an amino acid sequence having at least 72% identity with the amino acid sequence shown in SEQ ID NO: 42, specifically, 72% or more, preferably 75% or more, more preferably 80% or more, even more preferably 85% or more, even more preferably 90% or more, even more preferably 95% or more, even more preferably 96% or more, even more preferably 97% or more, even more preferably 98% or more, and even more preferably 99% or more identity. Amino acid sequences having at least 72% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0036】 Lipases comprising an amino acid sequence having at least 70% identity with the amino acid sequence shown in SEQ ID NO: 44 include lipases comprising an amino acid sequence having at least 70% identity with the amino acid sequence shown in SEQ ID NO: 44, specifically, 70% or more, preferably 75% or more, more preferably 80% or more, even more preferably 85% or more, even more preferably 90% or more, even more preferably 95% or more, even more preferably 96% or more, even more preferably 97% or more, even more preferably 98% or more, and even more preferably 99% or more. Amino acid sequences having at least 70% identity include amino acid sequences in which one or more amino acids are deleted, inserted, substituted, or added. 【0037】 "An amino acid sequence in which one or more amino acids are deleted, inserted, substituted, or added" includes an amino acid sequence in which one to 30 amino acids, preferably 20 or fewer, more preferably 10 or fewer, and even more preferably 5 or fewer amino acids are deleted, inserted, substituted, or added. 【0038】 Amino acid sequences having at least 79% identity with the amino acid sequence shown in SEQ ID NO: 14, amino acid sequences having at least 76% identity with the amino acid sequence shown in SEQ ID NO: 16, amino acid sequences having at least 81% identity with the amino acid sequence shown in SEQ ID NO: 18, amino acid sequences having at least 83% identity with the amino acid sequence shown in SEQ ID NO: 20, amino acid sequences having at least 96% identity with the amino acid sequence shown in SEQ ID NO: 22, amino acid sequences having at least 83% identity with the amino acid sequence shown in SEQ ID NO: 24, amino acid sequences having at least 90% identity with the amino acid sequence shown in SEQ ID NO: 26, amino acid sequences having at least 82% identity with the amino acid sequence shown in SEQ ID NO: 28, amino acid sequences having at least 73% identity with the amino acid sequence shown in SEQ ID NO: 30, and the sequence shown in SEQ ID NO: 32 Lipases comprising an amino acid sequence having at least 73% identity with the amino acid sequence, an amino acid sequence having at least 79% identity with the amino acid sequence shown in SEQ ID NO: 34, an amino acid sequence having at least 85% identity with the amino acid sequence shown in SEQ ID NO: 36, an amino acid sequence having at least 73% identity with the amino acid sequence shown in SEQ ID NO: 38, an amino acid sequence having at least 71% identity with the amino acid sequence shown in SEQ ID NO: 40, an amino acid sequence having at least 72% identity with the amino acid sequence shown in SEQ ID NO: 42, or an amino acid sequence having at least 70% identity with the amino acid sequence shown in SEQ ID NO: 44 include, for example, artificially created variants of lipases comprising the amino acid sequences shown in SEQ ID NOs: 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, or 44. Such mutants can be produced, for example, by introducing mutations into a gene encoding the amino acid sequence shown in SEQ ID NOs: 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, or 44 using known mutagenesis methods such as ultraviolet irradiation or site-directed mutagenesis, expressing the gene with the mutation, and selecting a protein with the desired lipase activity. Procedures for producing such mutants are well known to those skilled in the art. 【0039】 The lipase of the present invention has an amino acid sequence different from conventionally isolated or purified lipases and proteins predicted as triacylglycerol lipases in the NCBI protein sequence database. Examples of conventionally isolated or purified lipases include TLL (SEQ ID NO: 48), a lipase derived from Thermomyces lanuginosus; Lipr139 (SEQ ID NO: 2), a lipase derived from Cedecea sp.-16640 strain disclosed in Patent Document 1 as a lipase suitable for washing; Proteus bacterial lipase (hereinafter referred to as PvLip, SEQ ID NO: 4), disclosed in Patent Document 2 as the parent enzyme of a group of lipase variants suitable for washing; and Lipr138 (SEQ ID NO: 46), a metagenomic lipase disclosed in Patent Document 3 as a lipase suitable for washing. Furthermore, proteins predicted to be triacylglycerol lipases in the NCBI protein sequence database include the protein with accession number WP_123598507.1 (hereinafter referred to as PfLip, SEQ ID NO: 6), the protein with accession number WP_115457195.1 (hereinafter referred to as EtLip, SEQ ID NO: 8), the protein with accession number WP_135495634.1 (hereinafter referred to as EspLip, SEQ ID NO: 10), and the protein with accession number WP_005161363.1 (hereinafter referred to as YeLip, SEQ ID NO: 12). The enzymatic properties of these proteins registered in the database have not been reported to date. 【0040】 The lipases encoded by the amino acid sequences indicated by SEQ ID NOs: 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, or 44 are referred to as AncLip1, AncLip2, AncLip3, AncLip4, AncLip5, AncLip6, AncLip7, AncLip8, AncLip9, AncLip10, AncLip11, AncLip12, AncLip13, AncLip14, AncLip15, and AncLip16, respectively. AncLip1 shows 78% amino acid sequence identity with the lipase from Pseudomonas sp. registered in the NCBI protein sequence database with accession number WP_131062716.1. AncLip2 shows 75% amino acid sequence identity with the lipase from Proteus terrae registered in the NCBI protein sequence database with accession number WP_109394219.1. AncLip3 shows 80% amino acid sequence identity with the lipase from Enterobacillus tribolii registered in the NCBI protein sequence database with accession number WP_115457195.1. AncLip4 ​​shows 82% amino acid sequence identity with the lipase from Serratia registered in the NCBI protein sequence database with accession number WP_025122441.1. AncLip5 exhibits 95% amino acid sequence identity with the lipase from Yersinia aleksiciae registered in the NCBI protein sequence database with accession number WP_050126899.1. AncLip6 exhibits 82% amino acid sequence identity with the lipase from Enterobacteriaceae registered in the NCBI protein sequence database with accession number WP_045783583.1. AncLip7 exhibits 89% amino acid sequence identity with the lipase from Chania multitudinisentens registered in the NCBI protein sequence database with accession number WP_037407093.1. AncLip8 exhibits 81% amino acid sequence identity with the lipase from Serratia marcescens registered in the NCBI protein sequence database with accession number WP_033635162.1. AncLip9 exhibits 72% amino acid sequence identity with a lipase derived from Snodgrassella alvi, which is registered in the NCBI protein sequence database with accession number WP_100141403.1.AncLip10 shows 72% amino acid sequence identity with the lipase from Yersinia nurmii registered in the NCBI protein sequence database with accession number WP_049600386.1. AncLip11 shows 78% amino acid sequence identity with the lipase from Yersinia nurmii registered in the NCBI protein sequence database with accession number WP_049600386.1. AncLip12 shows 84% ​​amino acid sequence identity with the lipase from Yersinia enterocolitica registered in the NCBI protein sequence database with accession number WP_057631122.1. AncLip13 shows 72% amino acid sequence identity with the lipase from Snodgrassella registered in the NCBI protein sequence database with accession number WP_025329791.1. AncLip14 shows 70% amino acid sequence identity with the lipase from Serratia marcescens registered in the NCBI protein sequence database with accession number WP_033635162.1. AncLip15 shows 71% amino acid sequence identity with the lipase from Snodgrassella registered in the NCBI protein sequence database with accession number WP_025329791.1. AncLip16 shows 69% amino acid sequence identity with the lipase from Pseudomonas lundensis registered in the NCBI protein sequence database with accession number WP_097192271.1. 【0041】 In a preferred embodiment, the lipase of the present invention is a lipase comprising the amino acid sequence shown in SEQ ID NOs: 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, or 44, and more preferably a lipase comprising the amino acid sequence shown in SEQ ID NOs: 14, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, or 42. 【0042】 <2. Method for producing lipase> The lipase of the present invention can be produced, for example, by expressing the polynucleotide encoding the lipase of the present invention. Preferably, the lipase of the present invention can be produced from a transformant into which the polynucleotide encoding the lipase of the present invention has been introduced. For example, after obtaining a transformant by introducing the polynucleotide encoding the lipase of the present invention, or a vector containing it, into a host, the transformant can be cultured in a suitable medium to produce the lipase of the present invention from the polynucleotide encoding the lipase of the present invention introduced into the transformant. The lipase of the present invention can be obtained by isolating or purifying the produced lipase from the culture. 【0043】 The polynucleotide encoding the lipase of the present invention is an amino acid sequence shown in SEQ ID NO: 14 or an amino acid sequence having at least 79% identity thereto, an amino acid sequence shown in SEQ ID NO: 16 or an amino acid sequence having at least 76% identity thereto, an amino acid sequence shown in SEQ ID NO: 18 or an amino acid sequence having at least 81% identity thereto, an amino acid sequence shown in SEQ ID NO: 20 or an amino acid sequence having at least 83% identity thereto, an amino acid sequence shown in SEQ ID NO: 22 or an amino acid sequence having at least 96% identity thereto, an amino acid sequence shown in SEQ ID NO: 24 or an amino acid sequence having at least 83% identity thereto, an amino acid sequence shown in SEQ ID NO: 26 or an amino acid sequence having at least 90% identity thereto, an amino acid sequence shown in SEQ ID NO: 28 or an amino acid sequence having at least 82% identity thereto, sequence The present invention may be a polynucleotide encoding a lipase consisting of the amino acid sequence shown in sequence number 30 or an amino acid sequence having at least 73% identity thereto, the amino acid sequence shown in sequence number 32 or an amino acid sequence having at least 73% identity thereto, the amino acid sequence shown in sequence number 34 or an amino acid sequence having at least 79% identity thereto, the amino acid sequence shown in sequence number 36 or an amino acid sequence having at least 85% identity thereto, the amino acid sequence shown in sequence number 38 or an amino acid sequence having at least 73% identity thereto, the amino acid sequence shown in sequence number 40 or an amino acid sequence having at least 71% identity thereto, the amino acid sequence shown in sequence number 42 or an amino acid sequence having at least 72% identity thereto, or the amino acid sequence shown in sequence number 44 or an amino acid sequence having at least 70% identity thereto. Furthermore, the polynucleotide encoding the lipase of the present invention may be in the form of single-stranded or double-stranded DNA, RNA, or artificial nucleic acid, or it may be cDNA or chemically synthesized DNA that does not contain introns. 【0044】 The polynucleotide encoding the lipase of the present invention can be synthesized chemically or genetically based on the amino acid sequence of the lipase. For example, the polynucleotide can be chemically synthesized based on the amino acid sequence of the lipase of the present invention described above. For the chemical synthesis of the polynucleotide, nucleic acid synthesis contract services (e.g., those provided by Medical & Biological Laboratories, Inc., Genscript, etc.) can be used. Furthermore, the synthesized polynucleotide can be amplified by PCR, cloning, or other methods. 【0045】 Alternatively, the polynucleotide encoding the lipase of the present invention can be produced by introducing mutations into the polynucleotide synthesized by the above procedure using known mutagenesis methods such as ultraviolet irradiation or site-directed mutagenesis. For example, the polynucleotide encoding the lipase of the present invention can be obtained by introducing mutations into the polynucleotide of SEQ ID NOs. 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, or 43 using a known method, expressing the resulting polynucleotide and examining its lipase activity, and selecting the polynucleotide encoding a protein having the desired lipase activity. 【0046】 Site-directed mutagenesis into polynucleotides can be performed using any method, such as inverse PCR or annealing (Muramatsu et al., "Revised 4th Edition New Genetic Engineering Handbook," Yodosha, pp. 82-88). If necessary, various commercially available site-directed mutagenesis kits, such as Stratagene's QuickChange II Site-Directed Mutagenesis Kit or QuickChange Multi Site-Directed Mutagenesis Kit, can also be used. 【0047】 The polynucleotide encoding the lipase of the present invention can be incorporated into a vector. The type of vector containing the polynucleotide is not particularly limited and may be any vector such as plasmids, phages, phagemids, cosmids, viruses, YAC vectors, or shuttle vectors. The vector is, but is not limited, preferably a vector that can be amplified in bacteria, preferably in Bacillus bacteria (e.g., Bacillus subtilis or its mutants), and more preferably an expression vector that can induce the expression of the transgene in Bacillus bacteria. Among these, a shuttle vector, which can replicate in either Bacillus bacteria or other organisms, can be suitably used for recombinant production of the lipase of the present invention. Examples of preferred vectors include, but are not limited to, pHA3040SP64, pHSP64R, or pASP64 (Patent No. 3492935), shuttle vectors such as pHY300PLK (an expression vector capable of transforming both Escherichia coli and Bacillus subtilis; Jpn J Genet, 1985, 60:235-243), and pAC3 (Nucleic Acids Res, 1988, 16:8732); and plasmid vectors usable for transforming Bacillus bacteria such as pUB110 (J Bacteriol, 1978, 134:318-329) and pTA10607 (Plasmid, 1987, 18:8-15). Plasmid vectors derived from E. coli (e.g., pET22b(+), pBR322, pBR325, pUC57, pUC118, pUC119, pUC18, pUC19, pBluescript, etc.) can also be used. 【0048】 The above vector may include a DNA region containing a DNA replication initiation region or origin of replication. Alternatively, in the above vector, a regulatory sequence such as a promoter region, a terminator region, or a secretion signal region for secreting the expressed protein outside the cell may be operably ligated upstream of the polynucleotide encoding the lipase of the present invention (i.e., the lipase gene of the present invention). In this specification, "operably ligated" means that the gene and the regulatory region are arranged such that the gene can be expressed under the control of the regulatory region. 【0049】 The types of regulatory sequences such as the promoter region, terminator region, and secretion signal region are not particularly limited, and commonly used promoters and secretion signal sequences can be appropriately selected and used depending on the host to which the gene is introduced. For example, suitable examples of regulatory sequences that can be incorporated into a vector include the promoter and secretion signal sequence of the cellulase gene of Bacllus sp. KSM-S237 strain. 【0050】 Alternatively, the vector of the present invention may further incorporate marker genes (e.g., resistance genes for drugs such as ampicillin, neomycin, kanamycin, and chloramphenicol) for selecting a host into which the vector has been appropriately introduced. Alternatively, when using a nutrient-requiring strain as the host, a gene encoding an enzyme for synthesizing the required nutrients may be incorporated into the vector as a marker gene. Or, when using a selective medium that requires specific metabolism for growth, a gene related to that metabolism may be incorporated into the vector as a marker gene. An example of such a metabolism-related gene is the acetamidase gene for utilizing acetamide as a nitrogen source. 【0051】 The polynucleotide encoding the lipase of the present invention described above, along with the regulatory sequence and marker gene, can be linked by methods known in the art, such as SOE (splicing by overlap extension)-PCR (Gene, 1989, 77:61-68). The procedure for introducing the linked fragment into a vector is well known in the art. 【0052】 Transformed cells of the present invention can be obtained by introducing a vector containing a polynucleotide encoding the lipase of the present invention into a host, or by introducing a DNA fragment containing a polynucleotide encoding the lipase of the present invention into the host genome. 【0053】 Examples of host cells include microorganisms such as bacteria and filamentous fungi. Examples of bacteria include Escherichia coli, Staphylococcus, Enterococcus, Listeria, and Bacillus, among which Escherichia coli and Bacillus bacteria (for example, Bacillus subtilis Marburg No. 168 (Bacillus subtilis strain 168) or its mutants) are preferred. Examples of Bacillus subtilis mutants include the KA8AX protease 9-fold knockout strain described in J.Biosci.Bioeng.,2007,104(2):135-143, and the D8PA strain, which is an improved protein folding efficiency of the protease 8-fold knockout strain described in Biotechnol.Lett.,2011,33(9):1847-1852. Examples of filamentous fungi include the genera Trichoderma, Aspergillus, and Rizhopus. 【0054】 Methods commonly used in this field, such as protoplast synthesis and electroporation, can be used to introduce the vector into the host. By selecting strains in which the vector has been properly introduced based on indicators such as marker gene expression and nutritional requirements, the desired transformants into which the vector has been introduced can be obtained. 【0055】 Alternatively, a fragment comprising the polynucleotide encoding the lipase of the present invention, a regulatory sequence, and a marker gene can be directly introduced into the host genome. For example, a DNA fragment can be constructed by adding sequences complementary to the host genome to both ends of the linked fragment using a method such as SOE-PCR. This fragment can then be introduced into the host to induce homologous recombination between the host genome and the DNA fragment, thereby introducing the polynucleotide encoding the lipase of the present invention into the host genome. 【0056】 When a transformant into which the polynucleotide encoding the lipase of the present invention or a vector containing the same has been introduced is cultured in a suitable medium, the gene encoding the protein on the vector is expressed and the lipase of the present invention is produced. The medium used for culturing the transformant can be appropriately selected by a person skilled in the art according to the type of microorganism of the transformant. 【0057】 Alternatively, the lipase of the present invention may be expressed from a polynucleotide encoding the lipase of the present invention or its transcript using a cell-free translation system. A "cell-free translation system" is an in vitro transcription-translation system or in vitro translation system constructed by adding reagents such as amino acids necessary for protein translation to a suspension obtained by mechanically disrupting host cells. 【0058】 The lipase of the present invention, produced in the culture or cell-free translation system described above, can be isolated or purified by common methods used for protein purification, such as centrifugation, ammonium sulfate precipitation, gel chromatography, ion exchange chromatography, and affinity chromatography, either alone or in appropriate combinations. The protein recovered from the culture may be further purified by known means. 【0059】 <3. Detergent composition> The lipase obtained in this manner exhibits significantly higher resistance to inhibition of lipase activity by surfactants compared to known proteins, and shows significantly higher cleaning power even in the presence of surfactants. Here, "high resistance" means higher resistance compared to known proteins, specifically lipases consisting of amino acid sequences shown in SEQ ID NOs: 2, 4, 6, 8, 10, or 12, or proteins presumed to be lipases in the NCBI protein sequence database. Resistance to inhibition of lipase activity by surfactants can be evaluated using methods well known in the art. For example, a lipase solution and a surfactant solution are mixed, a solution of 4-nitrophenyl octanoate, a substrate of lipase, is added to the mixture, and the change in absorbance at 405 nm (OD / min) accompanying the release of 4-nitrophenol is measured. The difference ΔOD / min from the blank is determined as the lipase activity value (lipase activity value in the surfactant solution). Furthermore, the lipase activity value when a buffer is used instead of the surfactant solution (lipase activity value in the buffer) is determined, and the relative activity (%) is calculated by dividing the lipase activity value in the surfactant solution by the lipase activity value in the buffer and multiplying by 100. The higher the relative activity (%), the higher the resistance to inhibition of lipase activity by surfactants. The lipase of the present invention is a lipase with a relative lipase activity (%) of preferably 20% or more, more preferably 30% or more, and even more preferably 50% or more in a Triton X-100 solution (0.1% (w / v)) under the conditions of (3) in the example below. Furthermore, "high cleaning power" means a higher cleaning power compared to known proteins, specifically lipases consisting of amino acid sequences shown in SEQ ID NOs: 2, 4, 6, 8, 10, or 12, or proteins presumed to be lipases in the NCBI protein sequence database, for example, the ability to remove dirt in the washing or cleaning process. Cleaning power can be evaluated using methods well known in the art. For example, a cleaning solution containing lipase is added to a model stain containing a predetermined indicator substance (e.g., a highly soluble dye for fats such as Sudan III), and the cleaning process is performed under predetermined conditions. A portion of the cleaning solution is taken, and the concentration of the indicator substance in the model stain solubilized in the cleaning solution by the cleaning process is measured, for example, by absorbance measurement, and the difference from the blank can be determined as the cleaning power. 【0060】 The lipase of the present invention is useful as an enzyme for various detergent compositions, and is particularly useful as an enzyme for detergent compositions suitable for low-temperature washing. Here, "low temperature" includes temperatures below 40°C, below 35°C, below 30°C, and below 25°C, as well as temperatures above 5°C, above 10°C, and above 15°C. Additionally, temperatures between 5 and 40°C, 10 and 35°C, 15 and 30°C, and 15 and 25°C are also included. 【0061】 The amount of the lipase of the present invention incorporated into the detergent composition is not particularly limited as long as the amount of the lipase exhibits activity, but for example, it is preferably 0.1 mg or more, more preferably 1 mg or more, more preferably 5 mg or more per 1 kg of the detergent composition, and preferably 5000 mg or less, more preferably 1000 mg or less, and more preferably 500 mg or less. Furthermore, it is preferably 0.1 to 5000 mg, more preferably 1 to 1000 mg, and even more preferably 5 to 500 mg. 【0062】 The detergent composition preferably comprises a sulfosuccinate ester or a salt thereof in addition to the lipase of the present invention. Sulfosuccinate esters or salts thereof are known as components to be incorporated into detergent compositions (for example, Japanese Patent Application Publication No. 2019-182911). The sulfosuccinate ester or salt thereof is preferably a branched alkyl sulfosuccinate ester or salt thereof having a branched alkyl group having 9 to 12 carbon atoms, more preferably a branched alkyl sulfosuccinate ester or salt thereof having a branched alkyl group having 9 or 10 carbon atoms, and even more preferably a branched alkyl sulfosuccinate ester or salt thereof having a branched alkyl group having 10 carbon atoms. Furthermore, the sulfosuccinate ester or salt thereof is preferably a dibranched alkyl sulfosuccinate ester or salt thereof, wherein the two branched alkyl groups each have 9 to 12 carbon atoms; more preferably a dibranched alkyl sulfosuccinate ester or salt thereof, wherein the two branched alkyl groups each have 9 or 10 carbon atoms; even more preferably a dibranched alkyl sulfosuccinate ester or salt thereof, wherein the two branched alkyl groups each have 10 carbon atoms; and even more preferably bis-(2-propylheptyl)sulfosuccinate or a salt thereof. 【0063】 Examples of salts include alkali metal salts and alkanolamine salts, with alkali metal salts or alkanolamine salts being preferred, salts selected from sodium salts, potassium salts, triethanolamine salts, diethanolamine salts, and monoethanolamine salts being more preferred, and sodium salts being even more preferred. 【0064】 Examples of sulfosuccinate esters or their salts include compounds represented by the following formula 1. 【0065】 [ka] 【0066】 [In formula 1, R 1 , R 2is a branched alkyl group having 9 to 12 carbon atoms, respectively, and A 1 O, A 2 O are each an alkyleneoxy group having 2 to 4 carbon atoms, x1 and x2 are the average number of moles added, and are each a number of 0 or more and 10 or less, and M is a cation.〕 【0067】 In formula 1, R[[ENDEND]] 1 , R 2 are each preferably a branched alkyl group selected from a branched nonyl group, a branched decyl group, and a branched dodecyl group, and more preferably a branched decyl group. The branched decyl group is preferably a 2-propylheptyl group. 【0068】 In formula 1, A 1 O, A 2 O are each an alkyleneoxy group having 2 to 4 carbon atoms, and preferably an alkyleneoxy group having 2 or 3 carbon atoms from the viewpoint of lubricity with respect to water. In formula 1, x1 and x2 represent the average number of moles added of A 1 O, A 2 O, and are each a number of 0 or more and 10 or less, preferably 6 or less, more preferably 4 or less, still more preferably 2 or less, and even more preferably 0 from the viewpoint of lubricity with respect to water. 【0069】 In formula 1, M is a cation. M is preferably a cation other than a hydrogen ion. Examples of M include alkali metal ions such as lithium ion, sodium ion, and potassium ion, alkaline earth metal ions such as calcium ion and barium ion, organic ammonium ions such as triethanolammonium ion, diethanolammonium ion, monoethanolammonium ion, trimethylammonium ion, and monomethylammonium ion. M is preferably an alkali metal ion or an alkanolammonium ion from the viewpoint of dispersibility in water, more preferably sodium ion, potassium ion, triethanolammonium ion, diethanolammonium ion, or monoethanolammonium ion, and still more preferably sodium ion. 【0070】 The sulfosuccinate ester or salt thereof is preferably a compound represented by the following formula 1-1. The compound of formula 1-1 is a compound in which x1 and x2 in formula 1 are both 0. 【0071】 [ka] 【0072】 [In formula 1-1, R 1 , R 2 Each of these is a branched alkyl group having 9 to 12 carbon atoms, and M is a cation. R in Equation 1-1 1 , R 2 The specific and preferred examples of M are the same as in Equation 1. In a preferred embodiment, the sulfosuccinate ester or salt thereof is bis-(2-propylheptyl)sulfosuccinate or a salt thereof. 【0073】 The amount of sulfosuccinate ester or its salt blended into the detergent composition is preferably 0.01% by mass or more, more preferably 0.1% by mass or more, and preferably 2.0% by mass or less, and more preferably 1.0% by mass or less. It is also preferably 0.01 to 2.0% by mass, and even more preferably 0.1 to 1.0% by mass. 【0074】 The detergent composition may also contain various enzymes in addition to the lipase of the present invention. For example, hydrolases, oxidases, reductases, transferases, lyases, isomerases, ligases, synthetases, etc. Of these, lipases different from the lipase of the present invention, amylases, proteases, cellulases, keratinases, esterases, cutinases, pullulanases, pectinases, mannanases, glucosidases, glucanases, cholesterol oxidases, peroxidases, laccases, etc. are preferred, and proteases, cellulases, amylases, and lipases are particularly preferred. Examples of proteases include commercially available Alcalase, Esperase, Everlase, Savinase, Kannase, Progress Uno (registered trademark; Novozymes), PREFERENZ, EFFECTENZ, EXCELLENZ (registered trademark; DuPont), Lavergy (registered trademark; BASF), and KAP (Kao). Examples of cellulases include Celluclean, Carezyme (registered trademark; Novozymes), KAC, alkaline cellulase produced by Bacillus SP KSM-S237 strain described in Japanese Patent Publication No. 10-313859, and mutant alkaline cellulase described in Japanese Patent Publication No. 2003-313592 (all from Kao Corporation). Examples of amylases include Teramyl, Duramyl, Stainzyme, Stainzyme Plus, Amplify Prime (registered trademark; Novozymes), PREFERENZ, EFFECTENZ (registered trademark; DuPont), and KAM (Kao). Examples of lipases include Lipolase and Lipex (registered trademark; Novozymes). 【0075】 The detergent composition may contain known detergent components, and examples of such known detergent components include the following: 【0076】 (1) Surfactants The surfactant is incorporated into the detergent composition at a concentration of 0.5 to 60% by mass, and is preferably incorporated at a concentration of 10 to 45% by mass for powder detergent compositions and 20 to 90% by mass for liquid detergent compositions. Furthermore, when the detergent composition of the present invention is a laundry detergent or a detergent for automatic dishwashers, the surfactant is generally incorporated at a concentration of 1 to 10% by mass, preferably 1 to 5% by mass. 【0077】 Examples of surfactants used in the detergent composition include one or a combination of anionic surfactants, nonionic surfactants, amphoteric surfactants, and cationic surfactants, other than the sulfosuccinate esters or salts thereof mentioned above, but anionic surfactants and nonionic surfactants are preferred. 【0078】 As anionic surfactants, sulfate esters of alcohols having 10 to 18 carbon atoms, sulfate esters of alkoxylated alcohols having 8 to 20 carbon atoms, alkylbenzene sulfonates, paraffin sulfonates, α-olefin sulfonates, internal olefin sulfonates, α-sulfo fatty acid salts, α-sulfo fatty acid alkyl esters, or fatty acid salts are preferred. In the present invention, one or more anionic surfactants selected from linear alkylbenzene sulfonates having 10 to 14 carbon atoms in the alkyl chain, more preferably 12 to 14, and internal olefin sulfons having 12 to 20 carbon atoms in the alkylene chain, more preferably 16 to 18, are preferred, and as counterions, alkali metal salts and amines are preferred, with sodium and / or potassium, monoethanolamine, and diethanolamine being particularly preferred. For internal olefin sulfonic acid, see, for example, WO2017 / 098637. 【0079】 As nonionic surfactants, polyoxyalkylene alkyl (8-20 carbon atoms) ethers, alkyl polyglycosides, polyoxyalkylene alkyl (8-20 carbon atoms) phenyl ethers, polyoxyalkylene sorbitan fatty acid (8-22 carbon atoms) esters, polyoxyalkylene glycol fatty acid (8-22 carbon atoms) esters, and polyoxyethylene polyoxypropylene block polymers are preferred. In particular, as nonionic surfactants, polyoxyalkylene alkyl ethers obtained by adding 4-20 moles of alkylene oxide such as ethylene oxide or propylene oxide to an alcohol with 10-18 carbon atoms [such as having an HLB value (calculated by the Griffin method) of 10.5-15.0, preferably 11.0-14.5] are preferred. 【0080】 (2) Divalent metal ion scavenger The divalent metal ion scavenger is blended in an amount of 0.01 to 50% by mass, preferably 5 to 40% by mass. Examples of divalent metal ion scavengers used in the detergent composition of the present invention include condensed phosphates such as tripolyphosphate, pyrophosphate, and orthophosphate, aluminosilicates such as zeolites, synthetic layered crystalline silicates, nitrilotriacetate, ethylenediaminetetraacetate, citrate, isocitrate, and polyacetal carboxylate. Of these, crystalline aluminosilicate (synthetic zeolite) is particularly preferred, and among type A, type X, and type P zeolites, type A is particularly preferred. Synthetic zeolites with an average primary particle size of 0.1 to 10 μm, and particularly 0.1 to 5 μm, are preferably used. 【0081】 (3) Alkaline agents The alkaline agent is blended in an amount of 0.01 to 80% by mass, preferably 1 to 40% by mass. In the case of powder detergents, examples include alkali metal carbonates such as sodium carbonate, collectively known as dense ash or light ash, and amorphous alkali metal silicates such as JIS No. 1, No. 2, and No. 3. These inorganic alkaline agents are effective in forming the particle framework during detergent drying, resulting in a relatively hard detergent with excellent fluidity. Other alkalis include sodium sesquicarbonate and sodium bicarbonate, and phosphates such as tripolyphosphate also act as alkaline agents. In addition to the above alkaline agents, sodium hydroxide and mono, di, or triethanolamine can be used as alkaline agents in liquid detergents, and can also be used as counterions for surfactants. 【0082】 (4) Anti-redeposition agent The anti-redeposition agent is blended in an amount of 0.001 to 10% by mass, preferably 1 to 5% by mass. Examples of anti-redeposition agents used in the detergent composition of the present invention include polyethylene glycol, carboxylic acid polymers, polyvinyl alcohol, and polyvinylpyrrolidone. Of these, carboxylic acid polymers have the function of capturing metal ions in addition to their anti-redeposition ability, and the effect of dispersing solid particulate dirt from clothing into the washing bath. Carboxylic acid polymers are homopolymers or copolymers of acrylic acid, methacrylic acid, itaconic acid, etc., and copolymers obtained by copolymerizing the above monomers with maleic acid are preferred, with molecular weights of several thousand to 100,000 being preferred. In addition to the above carboxylic acid polymers, polymers such as polyglycidyl salts, cellulose derivatives such as carboxymethylcellulose, and aminocarboxylic acid polymers such as polyaspartic acid are also preferred because they have metal ion capturing, dispersing, and anti-redeposition abilities. 【0083】 (5) Bleach For example, it is preferable to include bleaching agents such as hydrogen peroxide and percarbonates in an amount of 1 to 10% by mass. When using bleaching agents, tetraacetylethylenediamine (TAED) or bleaching activators such as those described in Japanese Patent Publication No. 6-316700 can be included in an amount of 0.01 to 10% by mass. 【0084】 (6) Fluorescent agents Examples of fluorescent agents used in detergent compositions include biphenyl-type fluorescent agents (e.g., Chinopearl CBS-X) and stilbene-type fluorescent agents (e.g., DM-type fluorescent dyes). It is preferable to include 0.001 to 2% by mass of the fluorescent agent. 【0085】 (7) Other ingredients The detergent composition may contain builders, softeners, reducing agents (such as sulfites), antifoaming agents (such as silicones), fragrances, antibacterial and antifungal agents (such as Proxel [trade name], benzoic acid, etc.), and other additives known in the field of laundry detergents. 【0086】 The detergent composition can be manufactured by combining the lipase of the present invention obtained by the above method with the above known cleaning components according to conventional methods. The form of the detergent can be selected according to the application, for example, it can be a liquid, powder, granules, paste, solid, etc. 【0087】 The resulting detergent composition can be used as a laundry detergent, dishwashing detergent, bleach, hard surface cleaner, drain cleaner, denture cleaner, or disinfectant cleaner for medical instruments, but is more preferably used as a laundry detergent or dishwashing detergent, and more preferably as a laundry detergent (laundry detergent), a hand-washing dishwashing detergent, or a dishwashing machine detergent. Furthermore, the detergent composition is suitable for use at temperatures below 40°C, below 35°C, below 30°C, below 25°C, and above 5°C, above 10°C, and above 15°C. It is also suitable for use at temperatures between 5 and 40°C, 10 and 35°C, 15 and 30°C, and 15 and 25°C. Preferred uses include low-temperature (15-30°C) washing in laundry facilities and low-temperature (15-30°C) washing in automatic dishwashers. 【0088】 By using the cleaning agent composition of the present invention, it is possible to clean objects that require the removal of dirt (e.g., clothing, dishes, hard surfaces, drain pipes, dentures, medical instruments, etc.), that is, to remove dirt. Such a cleaning method involves bringing the object to be cleaned that requires the removal of dirt into contact with the cleaning agent composition of the present invention. Preferably, the dirt is sebum or dirt containing oils and fats derived from food. 【0089】 In the cleaning method of the present invention, to bring the object to be cleaned into contact with the cleaning agent composition, the object to be cleaned may be soaked in water in which the cleaning agent composition has been dissolved, or the cleaning agent composition may be applied directly to the object to be cleaned. In the method of the present invention, the object to be cleaned may be further washed by hand or in a washing machine after soaking or application of the cleaning agent composition, but this is not necessarily required. 【0090】 With regard to the embodiments described above, the present invention further discloses the following embodiments. <1> One of the following lipases: A lipase comprising the amino acid sequence shown in Sequence ID No. 14 or an amino acid sequence having at least 79% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 16 or an amino acid sequence having at least 76% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 18 or an amino acid sequence having at least 81% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 20 or an amino acid sequence having at least 83% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 22 or an amino acid sequence having at least 96% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 24 or an amino acid sequence having at least 83% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 26 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 28 or an amino acid sequence having at least 82% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 30 or an amino acid sequence having at least 73% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 32 or an amino acid sequence having at least 73% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 34 or an amino acid sequence having at least 79% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 36 or an amino acid sequence having at least 85% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 38 or an amino acid sequence having at least 73% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 40 or an amino acid sequence having at least 71% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 42 or an amino acid sequence having at least 72% identity thereto; and A lipase comprising the amino acid sequence shown in Sequence ID No. 44 or an amino acid sequence having at least 70% identity thereto. <2> The amino acid sequence consists of the sequence shown in SEQ ID NOs: 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, or 44. <1> The lipase described above. <3> <1> or <2> A polynucleotide encoding the lipase described above. <4> <3> A vector or DNA fragment containing the polynucleotides described above. <5> <4> Transformed cells containing the vector or DNA fragment described above. <6> It is a microorganism. <5> Transformed cells as described above. <7> It is either Escherichia coli or a bacterium of the genus Bacillus. <5> or <6> Transformed cells as described above. 【0091】 <8> <1> or <2> A detergent composition containing the lipase described above. <9> The product further contains a sulfosuccinate ester or a salt thereof, preferably a branched alkyl sulfosuccinate ester or a salt thereof having a branched alkyl group with 9 to 12 carbon atoms, more preferably a branched alkyl sulfosuccinate ester or a salt thereof having a branched alkyl group with 9 or 10 carbon atoms, and even more preferably a branched alkyl sulfosuccinate ester or a salt thereof having a branched alkyl group with 10 carbon atoms. <8> The cleaning agent composition described above. <10> A disulfosuccinate or salt thereof, preferably a dibranched alkyl sulfosuccinate or salt thereof in which two branched alkyl groups each have 9 to 12 carbon atoms, more preferably a dibranched alkyl sulfosuccinate or salt thereof in which two branched alkyl groups each have 9 or 10 carbon atoms, even more preferably a dibranched alkyl sulfosuccinate or salt thereof in which two branched alkyl groups each have 10 carbon atoms, and even more preferably bis-(2-propylheptyl)sulfosuccinate or salt thereof. <8> The cleaning agent composition described above. <11> It is a laundry detergent or a dishwashing detergent. <8> ~ <10> A detergent composition as described in any of the following. <12> It is either a powder or a liquid. <8> ~ <11> A detergent composition as described in any of the following. <13> Used at low temperatures, <8> ~ <12> A detergent composition as described in any of the following. <14> Used at temperatures below 40℃, below 35℃, below 30℃, below 25℃, and above 5℃, above 10℃, above 15℃, or used at temperatures between 5 and 40℃, 10 and 35℃, 15 and 30℃, and 15 and 25℃. <13> The cleaning agent composition described above. 【0092】 <15> <8> ~ <14> A method for cleaning dirt using a cleaning agent composition described in any of the above. <16> The object to be cleaned and <8> ~ <14> This includes contacting the cleaning agent composition described in any of the above, <15> Methods used. <17> The stain is sebum or contains oils and fats derived from food. <15> or <16> Methods used. 【0093】 <18> Use of any of the following lipases for the manufacture of detergent compositions: A lipase comprising the amino acid sequence shown in Sequence ID No. 14 or an amino acid sequence having at least 79% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 16 or an amino acid sequence having at least 76% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 18 or an amino acid sequence having at least 81% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 20 or an amino acid sequence having at least 83% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 22 or an amino acid sequence having at least 96% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 24 or an amino acid sequence having at least 83% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 26 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 28 or an amino acid sequence having at least 82% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 30 or an amino acid sequence having at least 73% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 32 or an amino acid sequence having at least 73% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 34 or an amino acid sequence having at least 79% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 36 or an amino acid sequence having at least 85% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 38 or an amino acid sequence having at least 73% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 40 or an amino acid sequence having at least 71% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 42 or an amino acid sequence having at least 72% identity thereto; and A lipase comprising the amino acid sequence shown in Sequence ID No. 44 or an amino acid sequence having at least 70% identity thereto. <19> The lipase is a lipase consisting of the amino acid sequence shown in SEQ ID NOs: 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, or 44. <18> Use as described above. <20> The detergent composition is a laundry detergent or a dishwashing detergent. <18> or <19> Use as described above. <21> The cleaning agent composition is in powder or liquid form. <18> ~ <20> Use as described in any of the following. <22> The detergent composition is intended for use at low temperatures. <18> ~ <21> Use as described in any of the following. <23> The detergent composition is used at temperatures of 40°C or below, 35°C or below, 30°C or below, 25°C or below, and 5°C or above, 10°C or above, 15°C or above, or at temperatures of 5-40°C, 10-35°C, 15-30°C, 15-25°C. <22> Use as described above. 【0094】 <24> Use of one of the following lipases for cleaning stains: A lipase comprising the amino acid sequence shown in Sequence ID No. 14 or an amino acid sequence having at least 79% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 16 or an amino acid sequence having at least 76% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 18 or an amino acid sequence having at least 81% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 20 or an amino acid sequence having at least 83% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 22 or an amino acid sequence having at least 96% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 24 or an amino acid sequence having at least 83% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 26 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 28 or an amino acid sequence having at least 82% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 30 or an amino acid sequence having at least 73% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 32 or an amino acid sequence having at least 73% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 34 or an amino acid sequence having at least 79% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 36 or an amino acid sequence having at least 85% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 38 or an amino acid sequence having at least 73% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 40 or an amino acid sequence having at least 71% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 42 or an amino acid sequence having at least 72% identity thereto; and A lipase comprising the amino acid sequence shown in Sequence ID No. 44 or an amino acid sequence having at least 70% identity thereto. <25> The lipase is a lipase consisting of the amino acid sequence shown in SEQ ID NOs: 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, or 44. <24> Use as described above. <26> The stain is sebum or contains oils and fats derived from food. <24> or <25> Use as described above. <27> The washing is done at a low temperature. <24> ~ <26> Use as described in any of the following. <28> The washing is performed at temperatures below 40°C, below 35°C, below 30°C, below 25°C, and above 5°C, above 10°C, and above 15°C, or at temperatures between 5 and 40°C, 10 and 35°C, 15 and 30°C, and 15 and 25°C. <27> Use as described above. [Examples] 【0095】 The present invention will be described in more detail below based on examples, but the present invention is not limited thereto. 【0096】 (1) Construction of lipase expression plasmid The lipase expression plasmid was constructed using the VHH expression plasmid of Sequence ID No. 26, which contains a Bacillus subtilis spoVG gene-derived promoter as described in WO2021 / 153129, as a template. The plasmid was then constructed by replacing each lipase gene with the full-length ORF containing the VHH gene of the above plasmid via an in-fusion reaction. The artificially synthesized lipase genes were Lipr139, PvLip, PfLip, EtLip, EspLip, YeLip, AncLip1, AncLip2, AncLip3, AncLip4, AncLip5, AncLip6, AncLip7, AncLip8, AncLip9, AncLip10, AncLip11, AncLip12, AncLip13, and A. ncLip14, AncLip15, AncLip16, Lipr138 (polynucleotides of SEQ ID NOs. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43 and 45 respectively, and SEQ ID NOs. 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30 respectively) Plasmids pHY-Lipr139, pHY-PvLip, pHY-PfLip, pHY-EtLip, pHY-EspLip, pHY-YeLip, pHY-AncLip1, pHY-AncLip2, pHY-AncLip3, pHY-AncLip4, pHY-AncLip5, pHY-AncLip6, pHY-AncLip7, pHY-AncLip8, pHY-AncLip9, pHY-AncLip10, pHY-AncLip11, pHY-AncLip12, pHY-AncLip13, pHY-AncLip14, pHY-AncLip15, pHY-AncLip16, and pHY-Lipr138 were constructed from the amino acid sequences (32, 34, 36, 38, 40, 42, 44, and 46), respectively. Furthermore, the amino acid sequences represented by sequence numbers 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, and 44 are novel lipase sequences that are presumed to have been present in the ancestral organisms of the existing lipase-producing bacteria belonging to the Proteus / Yersinia clade.pHY-amyEsig-TLL was constructed by in-fusion reaction, replacing the full-length ORF containing the Lipr139 gene of plasmid pHY-Lipr139 with an artificially synthesized lipase TLL gene (encoding the polynucleotide of SEQ ID NO: 47 and the amino acid sequence of SEQ ID NO: 48) in which a signal sequence derived from Bacillus subtilis amyE (encoding the polynucleotide of SEQ ID NO: 49 and the amino acid sequence of SEQ ID NO: 50) was ligated to the N-terminus. 【0097】 (2) Preparation of lipase solution Lipase expression plasmids were introduced into Bacillus subtilis strains by protoplast, and the cultures were incubated in 2×L-maltose medium (2% tryptone, 1% yeast extract, 1% NaCl, 7.5% maltose, 7.5 ppm manganese sulfate pentahydrate, 0.04% calcium chloride dihydrate, 15 ppm tetracycline; % is (w / v)%) at 30°C for 2 days. The culture supernatant containing lipase was then collected by centrifugation. The culture supernatant was buffered with 10 mM Tris-HCl + 0.01% Triton-X100 (pH 7.0) by dialysis, and the lipase concentration was quantified from the band intensity of SDS-PAGE. 【0098】 (3) Activity measurement in surfactant solution 4-nitrophenyl octanoate (SIGMA) was used as the substrate. Lipase activity can be determined by measuring the rate of increase in absorbance associated with the release of 4-nitrophenol by the action of lipase. 20 mM 4-nitrophenyl octanoate in 20 mM Tris-HCl (pH 7.0) was used as the substrate solution. 0.1% (w / v) of SDS or Triton X-100 added to 20 mM Tris-HCl (pH 7.0) was used as the surfactant solution. 5 μL of lipase solution adjusted to 2 ppm and 100 μL of surfactant solution were mixed in each well of a 96-well assay plate, and 10 μL of substrate solution was added. The absorbance change (OD / min) at 405 nm was measured at 30°C. The difference ΔOD / min from the blank (sample without enzyme addition) was taken as the activity value. For each lipase, the relative activity (%) was calculated by dividing the activity value in the surfactant solution by the activity value in the buffer (activity in 20 mM Tris-HCl (pH 7.0) instead of the surfactant solution) and multiplying by 100 (Figure 1). Lipr139, PvLip, and Lipr138, whose suitability for washing is disclosed in TLL, Patent Documents 1, 2, and 3, as well as naturally occurring lipase sequences PfLip, EtLip, EspLip, and YeLip, were significantly inhibited in activity in the presence of SDS and Triton X-100. In comparison, the novel ancestral lipases AncLip1, AncLip2, AncLip3, AncLip4, AncLip5, AncLip6, AncLip7, AncLip8, AncLip9, AncLip10, AncLip11, AncLip12, AncLip13, AncLip14, AncLip15, and AncLip16 showed high resistance to activity inhibition by Triton X-100. Furthermore, AncLip3, AncLip4, AncLip6, AncLip8, AncLip9, AncLip12, and AncLip13 showed high resistance to activity inhibition by SDS. 【0099】 (4) Activity measurement in model washing solution 20 mM 4-nitrophenyl octanoate in 20 mM Tris-HCl (pH 7.0) was used as the substrate solution. The aqueous media shown in Table 1 were used as the model washing solution. 2 μL of lipase solution adjusted to 4 ppm and 100 μL of the test solution (model washing solution or 20 mM Tris-HCl (pH 7.0)) were mixed in each well of a 96-well assay plate, and 10 μL of the substrate solution was added. The absorbance change at 405 nm (OD / min) was measured at 30°C. The difference ΔOD / min from the blank (sample without enzyme addition) was calculated. 10 μL of 20 mM Tris-HCl (pH 7.0) containing 31-500 μM 4-nitrophenol was mixed with 100 μL of each test solution, and the absorbance at 405 nm was measured to create a calibration curve. The activity value was calculated from the calibration curve and ΔOD / min value of each test solution, determining the release rate of 4-nitrophenol per minute (μM / min). For each lipase, the relative activity (%) was calculated by dividing the activity value when using the model washing solution as the test solution by the activity value (activity in buffer) when using 20 mM Tris-HCl (pH 7.0) as the test solution, and multiplying by 100 (Figure 2). Lipr139, PvLip, and Lipr138, whose suitability for washing is disclosed in TLL and Patent Documents 1, 2, and 3, as well as the naturally occurring lipase sequences PfLip, EtLip, EspLip, and YeLip, were significantly inhibited in activity in the model washing solution. In comparison, the novel ancestral lipases AncLip1, AncLip3, AncLip4, AncLip6, AncLip7, AncLip8, AncLip9, AncLip10, AncLip11, AncLip12, AncLip13, AncLip14, AncLip15, and AncLip16 showed high resistance to activity inhibition in the model washing solution. 【0100】 [Table 1] 【0101】 (5) Cleaning evaluation in model cleaning solution Beef tallow (SIGMA, 03-0660) and rapeseed oil (SIGMA, 23-0450) were mixed in a weight ratio of 9:1, dissolved in three times the volume of chloroform, and then colored with 0.2 wt% Sudan III to create the model stain. 10 μL of the model stain was dropped onto the bottom of each well of a 96-well polypropylene deep-well plate, and the chloroform was evaporated and dried to create the stain plate. The cleaning solution was prepared by adding 1 / 100 volume of 240 ppm lipase solution to the model cleaning solution listed in Table 1. 300 μL of the cleaning solution was slowly added to each soiled plate, and immersion cleaning was performed by letting it stand at room temperature (approximately 22°C) for 15 minutes. 100 μL of the cleaning solution was carefully removed from each plate, taking care not to touch the soil at the bottom, and transferred to a new 96-well plate. To quantify Sudan III in the model soil that was solubilized in the cleaning solution by immersion cleaning, the absorbance at 500 nm (A500) was measured. The value ΔA500, obtained by subtracting the pre-cleaning A500 from the post-cleaning A500, corresponds to the amount of oil released into the cleaning solution and can be used as an indicator of cleaning power. The cleaning power ΔA500 for each lipase is shown in Figure 3. Lipr139, whose suitability for cleaning is disclosed in Patent Document 1, showed higher cleaning power in the model cleaning solution compared to TLL, Lipr138, PvLip, PfLip, EtLip, EspLip, and YeLip. On the other hand, the novel ancestral lipases AncLip1, AncLip3, AncLip4, AncLip5, AncLip6, AncLip7, AncLip8, AncLip9, AncLip10, AncLip11, AncLip12, AncLip13, AncLip14, and AncLip15 showed significantly higher cleaning power in the model cleaning solution compared to Lipr139. 【0102】 Phylogenetic trees were constructed using the amino acid sequences of Lipr139, PvLip, PfLip, EtLip, EspLip, YeLip, AncLip1, AncLip2, AncLip3, AncLip4, AncLip5, AncLip6, AncLip7, AncLip8, and AncLip9 (sequence numbers 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, and 30, respectively). Multiple alignment was performed using clustalW with Genetyx, and the phylogenetic trees were constructed using the neighbor-joining method (NJ method) (Figure 4).

Claims

[Claim 1] Any of the following lipases that exhibits higher resistance to inhibition of lipase activity in an aqueous solution containing 0.2% by mass of sodium bis-(2-propylheptyl)sulfosuccinate, 2% by mass of lauryl(2-hydroxy-3-sulfopropyl)dimethylbetaine and 6.5% by mass of butyl diglycol compared to the lipase comprising the amino acid sequence shown in Sequence ID No. 46: A lipase comprising the amino acid sequence shown in Sequence ID No. 32 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 14 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 18 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 20 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 24 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 26 or an amino acid sequence having at least 95% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 28 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 30 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 34 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 36 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 38 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 40 or an amino acid sequence having at least 90% identity thereto; A lipase comprising the amino acid sequence shown in Sequence ID No. 42 or an amino acid sequence having at least 90% identity thereto; and A lipase comprising the amino acid sequence shown in Sequence ID No. 44 or an amino acid sequence having at least 90% identity thereto. [Claim 2] A polynucleotide encoding the lipase described in claim 1. [Claim 3] A vector or DNA fragment comprising the polynucleotide described in claim 2. [Claim 4] Transformed cells containing the vector or DNA fragment described in claim 3. [Claim 5] A transformed cell according to claim 4, which is a microorganism. [Claim 6] A detergent composition containing the lipase described in claim 1. [Claim 7] The detergent composition according to claim 6, which is a laundry detergent or a dishwashing detergent. [Claim 8] The cleaning agent composition according to claim 7, which is a powder or a liquid. [Claim 9] A detergent composition according to claim 7 or 8, for use at low temperatures. [Claim 10] The detergent composition according to claim 9, which is used at a temperature of 5 to 40°C. [Claim 11] A method for cleaning dirt using the cleaning agent composition described in claim 6.

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