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Article(id=1226236833328181880, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1226236828399878330, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20240718, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1731427200000, receivedDateStr=2024-11-13, revisedDate=null, revisedDateStr=null, acceptedDate=1740067200000, acceptedDateStr=2025-02-21, onlineDate=1770287243444, onlineDateStr=2026-02-05, pubDate=1746288000000, pubDateStr=2025-05-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1770287243444, onlineIssueDateStr=2026-02-05, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1770287243444, creator=13701087609, updateTime=1770287243444, updator=13701087609, issue=Issue{id=1226236828399878330, tenantId=1146029695717560320, journalId=1192105938417971205, year='2025', volume='65', issue='5', pageStart='1831', pageEnd='2319', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1770287242269, creator=13701087609, updateTime=1770344517883, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1226477059812274835, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1226236828399878330, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1226477059816469140, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1226236828399878330, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=2201, endPage=2213, ext={EN=ArticleExt(id=1226236834150265492, articleId=1226236833328181880, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Heterologous expression and enzymatic characterization of a thermostable and alkaline-stable esterase from Streptomyces griseus, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

[Objective] This study characterized a novel esterase EstE from Streptomyces griseus by heterologous expression in Escherichia coli and systematically evaluates its thermostability, alkaline stability, and the effects of various additives (metal ions, detergents, and organic solvents) on its enzymatic activity to explore its potential for industrial applications. [Methods] We synthesized the gene estE' encoding the same amino acid sequence as the native gene by optimizing the original sequence of estE from S. griseus. We then constructed the recombinant plasmid carrying the optimized gene by ligating the gene into the pET-28b(+) vector. The esterase EstE was then expressed under the induction of IPTG and purified via Co2+ affinity chromatography. Furthermore, the enzymatic properties of the purified EstE were determined by the p-nitrophenol method, and bioinformatics analysis was performed for this enzyme. [Results] EstE consisted of 289 amino acid residues, with a molecular weight of 31.6 kDa. It belonged to the GDS(L) family, with Ser16, Asp194, and His224 forming its catalytic triad. The enzyme showed the optimal activity at 40 ℃ and pH 8.5, with the highest catalytic efficiency (specific activity of 61.03 U/mg) observed in the case of p-nitrophenyl acetate as a substrate. EstE demonstrated robust thermostability, with the relative activity of 50% after 156.11 h of incubation at 40 ℃ and 2.67 h of incubation at 100 ℃. Moreover, it showed excellent alkaline stability, with the relative activity exceeding 80% after incubation at pH 8.5 for 100 h. In addition, this enzyme exhibited excellent tolerance to organic solvents, maintaining stable activity in the presence of 30% DMSO. [Conclusion] A novel esterase EstE from S. griseus is successfully obtained through heterologous expression, demonstrating excellent catalytic properties, thermostability, alkaline stability, and organic solvent tolerance, positioning it as a promising candidate for industrial applications.

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【目的】通过在大肠杆菌中异源表达，获得一种来源于灰色链霉菌(Streptomyces griseus)的新型酯酶EstE，并系统评估其热稳定性、碱稳定性以及不同添加剂(金属离子、去垢剂和有机溶剂)对酶活性的影响，以探索其在工业应用中的潜力。【方法】对灰色链霉菌中酯酶基因estE进行人工优化，合成编码相同氨基酸序列的酯酶基因estE'，将其与载体pET-28b(+)连接后，构建含该酯酶基因的重组质粒。通过IPTG诱导表达，采用Co2+亲和层析纯化得到酯酶EstE，随后利用对硝基苯酚法对该酶的酶学性质进行鉴定，并对其进行生物信息学分析。【结果】EstE由289个氨基酸组成，分子量约为31.6 kDa，属于GDS(L)家族，其催化三联体由Ser16、Asp194、His224构成。该酶的最适温度和最适pH分别为40 ℃和pH 8.5，以对硝基苯酚乙酸酯为底物时催化活性最高，比酶活为61.03 U/mg。在热稳定性方面，酯酶EstE在40 ℃和100 ℃条件下的半衰期分别为156.11 h和2.67 h；在碱稳定性方面，其在pH 8.5下孵育100 h后，仍能维持80%以上的相对酶活性。在有机溶剂耐受性方面，当存在30% DMSO时，其酶活基本不变。【结论】通过异源表达，成功获得了一种来源于灰色链霉菌的新型酯酶EstE。该酶具有优异的催化特性、热稳定性、碱稳定性和有机溶剂耐受性，为其在工业领域中的应用奠定了基础。

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作者贡献声明

李全发：实验设计、数据收集和处理、论文撰写和修改；房金鑫：数据收集和处理、论文撰写和修改；喻娇：数据收集；陈菁菁：协助实验操作；王宝娟：研究构思和设计、论文撰写和修改。

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Microbial carboxyl esterases: classification, properties and application in biocatalysis[J]. FEMS Microbiology Reviews, 2002, 26(1): 73-81., articleTitle=Microbial carboxyl esterases: classification, properties and application in biocatalysis, refAbstract=null), Reference(id=1226592770303898409, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2020, volume=170, issue=null, pageStart=104704, pageEnd=null, url=null, language=null, rfNumber=[2], rfOrder=1, authorNames=WANG BJ, WU S, CHANG X, CHEN J, MA JX, WANG P, ZHU GP, journalName=Pesticide Biochemistry and Physiology, refType=null, unstructuredReference=WANG BJ, WU S, CHANG X, CHEN J, MA JX, WANG P, ZHU GP. Characterization of a novel hyper-thermostable and chlorpyrifos-hydrolyzing carboxylesterase EstC: a representative of the new esterase family XIX[J]. Pesticide Biochemistry and Physiology, 2020, 170: 104704., articleTitle=Characterization of a novel hyper-thermostable and chlorpyrifos-hydrolyzing carboxylesterase EstC: a representative of the new esterase family XIX, refAbstract=null), Reference(id=1226592770417144624, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2021, volume=205, issue=null, pageStart=111882, pageEnd=null, url=null, language=null, rfNumber=[3], rfOrder=2, authorNames=JOHAN UUM, RAHMAN RNZRA, KAMARUDIN NHA, ALI MSM, journalName=Colloids and Surfaces B: Biointerfaces, refType=null, unstructuredReference=JOHAN UUM, RAHMAN RNZRA, KAMARUDIN NHA, ALI MSM. An integrated overview of bacterial carboxylesterase: structure, function and biocatalytic applications[J]. Colloids and Surfaces B: Biointerfaces, 2021, 205: 111882., articleTitle=An integrated overview of bacterial carboxylesterase: structure, function and biocatalytic applications, refAbstract=null), Reference(id=1226592770584916792, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2020, volume=19, issue=1, pageStart=169, pageEnd=null, url=null, language=null, rfNumber=[4], rfOrder=3, authorNames=CHANDRA P, ENESPA, SINGH R, ARORA PK, journalName=Microbial Cell Factories, refType=null, unstructuredReference=CHANDRA P, ENESPA, SINGH R, ARORA PK. Microbial lipases and their industrial applications: a comprehensive review[J]. Microbial Cell Factories, 2020, 19(1): 169., articleTitle=Microbial lipases and their industrial applications: a comprehensive review, refAbstract=null), Reference(id=1226592770702357308, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=1999, volume=343, issue=null, pageStart=177, pageEnd=183, url=null, language=null, rfNumber=[5], rfOrder=4, authorNames=ARPIGNY JL, JAEGER KE, journalName=Biochemical Journal, refType=null, unstructuredReference=ARPIGNY JL, JAEGER KE. Bacterial lipolytic enzymes: classification and properties[J]. Biochemical Journal, 1999, 343(Pt 1): 177-183., articleTitle=Bacterial lipolytic enzymes: classification and properties, refAbstract=null), Reference(id=1226592770803020609, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2022, volume=13, issue=null, pageStart=855658, pageEnd=null, url=null, language=null, rfNumber=[6], rfOrder=5, authorNames=LIU XY, ZHOU MY, SUN R, XING S, WU T, HE HL, CHEN JB, BIELICKI JK, journalName=Frontiers in Microbiology, refType=null, unstructuredReference=LIU XY, ZHOU MY, SUN R, XING S, WU T, HE HL, CHEN JB, BIELICKI JK. Characterization of a novel esterase Est33 from an antarctic bacterium: a representative of a new esterase family[J]. Frontiers in Microbiology, 2022, 13: 855658., articleTitle=Characterization of a novel esterase Est33 from an antarctic bacterium: a representative of a new esterase family, refAbstract=null), Reference(id=1226592770920461126, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2019, volume=null, issue=null, pageStart=1, pageEnd=35, url=null, language=null, rfNumber=[7], rfOrder=6, authorNames=KOVACIC F, BABIC N, KRAUSS U, JAEGER KE, journalName=Aerobic Utilization of Hydrocarbons, Oils and Lipids, refType=null, unstructuredReference=KOVACIC F, BABIC N, KRAUSS U, JAEGER KE. Classification of lipolytic enzymes from bacteria[M]//ROJO F eds. Aerobic Utilization of Hydrocarbons, Oils and Lipids. Handbook of Hydrocarbon and Lipid Microbiology. Cham: Springer International Publishing, 2019: 1-35., articleTitle=Classification of lipolytic enzymes from bacteria, refAbstract=null), Reference(id=1226592771046290251, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2023, volume=63, issue=14, pageStart=2057, pageEnd=2073, url=null, language=null, rfNumber=[8], rfOrder=7, authorNames=WU H, CHEN QM, ZHANG WL, MU WM, journalName=Critical Reviews in Food Science and Nutrition, refType=null, unstructuredReference=WU H, CHEN QM, ZHANG WL, MU WM. Overview of strategies for developing high thermostability industrial enzymes: discovery, mechanism, modification and challenges[J]. Critical Reviews in Food Science and Nutrition, 2023, 63(14): 2057-2073., articleTitle=Overview of strategies for developing high thermostability industrial enzymes: discovery, mechanism, modification and challenges, refAbstract=null), Reference(id=1226592771184702297, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2023, volume=425, issue=null, pageStart=138915, pageEnd=null, url=null, language=null, rfNumber=[9], rfOrder=8, authorNames=SUJITHA P, SHANTHI C, journalName=Journal of Cleaner Production, refType=null, unstructuredReference=SUJITHA P, SHANTHI C. Importance of enzyme specificity and stability for the application of proteases in greener industrial processing-a review[J]. Journal of Cleaner Production, 2023, 425: 138915., articleTitle=Importance of enzyme specificity and stability for the application of proteases in greener industrial processing-a review, refAbstract=null), Reference(id=1226592771289559898, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2023, volume=290, issue=9, pageStart=2394, pageEnd=2411, url=null, language=null, rfNumber=[10], rfOrder=9, authorNames=MARCHETTI A, ORLANDO M, MANGIAGALLI M, LOTTI M, journalName=The FEBS Journal, refType=null, unstructuredReference=MARCHETTI A, ORLANDO M, MANGIAGALLI M, LOTTI M. A cold-active esterase enhances mesophilic properties through Mn2+ binding[J]. The FEBS Journal, 2023, 290(9): 2394-2411., articleTitle=A cold-active esterase enhances mesophilic properties through Mn2+ binding, refAbstract=null), Reference(id=1226592771407000416, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2024, volume=138, issue=3, pageStart=181, pageEnd=187, url=null, language=null, rfNumber=[11], rfOrder=10, authorNames=FANG JX, AN LH, YU J, MA JX, ZHOU RJ, WANG BJ, journalName=Journal of Bioscience and Bioengineering, refType=null, unstructuredReference=FANG JX, AN LH, YU J, MA JX, ZHOU RJ, WANG BJ. Characterization of a novel carboxylesterase from Streptomyces lividans TK24 and site-directed mutagenesis for its thermostability[J]. Journal of Bioscience and Bioengineering, 2024, 138(3): 181-187., articleTitle=Characterization of a novel carboxylesterase from Streptomyces lividans TK24 and site-directed mutagenesis for its thermostability, refAbstract=null), Reference(id=1226592772786926439, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2016, volume=51, issue=10, pageStart=1434, pageEnd=1444, url=null, language=null, rfNumber=[12], rfOrder=11, authorNames=KUMAR A, SHARMA M, BHARDWAJ PK, VATS SK, SINGH D, KUMAR S, journalName=Process Biochemistry, refType=null, unstructuredReference=KUMAR A, SHARMA M, BHARDWAJ PK, VATS SK, SINGH D, KUMAR S. Copper, zinc superoxide dismutase from Caragana jubata: a thermostable enzyme that functions under a broad pH and temperature window[J]. Process Biochemistry, 2016, 51(10): 1434-1444., articleTitle=Copper, zinc superoxide dismutase from Caragana jubata: a thermostable enzyme that functions under a broad pH and temperature window, refAbstract=null), Reference(id=1226592772875006828, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2004, volume=43, issue=6, pageStart=534, pageEnd=552, url=null, language=null, rfNumber=[13], rfOrder=12, authorNames=AKOH CC, LEE GC, LIAW YC, HUANG TH, SHAW JF, journalName=Progress in Lipid Research, refType=null, unstructuredReference=AKOH CC, LEE GC, LIAW YC, HUANG TH, SHAW JF. GDSL family of serine esterases/lipases[J]. Progress in Lipid Research, 2004, 43(6): 534-552., articleTitle=GDSL family of serine esterases/lipases, refAbstract=null), Reference(id=1226592773072139121, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2021, volume=25, issue=2, pageStart=115, pageEnd=128, url=null, language=null, rfNumber=[14], rfOrder=13, authorNames=CHANG X, WU S, CHEN J, XIONG SQ, WANG P, SHI XQ, WANG A, WANG BJ, journalName=Extremophiles, refType=null, unstructuredReference=CHANG X, WU S, CHEN J, XIONG SQ, WANG P, SHI XQ, WANG A, WANG BJ. Characterization of a carboxylesterase with hyper-thermostability and alkali-stability from Streptomyces lividans TK24[J]. Extremophiles, 2021, 25(2): 115-128., articleTitle=Characterization of a carboxylesterase with hyper-thermostability and alkali-stability from Streptomyces lividans TK24, refAbstract=null), Reference(id=1226592773189579640, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2001, volume=309, issue=1, pageStart=215, pageEnd=226, url=null, language=null, rfNumber=[15], rfOrder=14, authorNames=VAN POUDEROYEN G, EGGERT T, JAEGER KE, DIJKSTRA BW, journalName=Journal of Molecular Biology, refType=null, unstructuredReference=VAN POUDEROYEN G, EGGERT T, JAEGER KE, DIJKSTRA BW. The crystal structure of Bacillus subtilis lipase: a minimal α/β hydrolase fold enzyme[J]. Journal of Molecular Biology, 2001, 309(1): 215-226., articleTitle=The crystal structure of Bacillus subtilis lipase: a minimal α/β hydrolase fold enzyme, refAbstract=null), Reference(id=1226592773277660026, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2022, volume=52, issue=4, pageStart=394, pageEnd=403, url=null, language=null, rfNumber=[16], rfOrder=15, authorNames=KNEPP ZJ, GHANER A, ROOT KT, journalName=Preparative Biochemistry & Biotechnology, refType=null, unstructuredReference=KNEPP ZJ, GHANER A, ROOT KT. Purification and refolding protocol for cold-active recombinant esterase Aa SGNH1 from Aphanizomenon flos-aquae expressed as insoluble inclusion bodies[J]. Preparative Biochemistry & Biotechnology, 2022, 52(4): 394-403., articleTitle=Purification and refolding protocol for cold-active recombinant esterase Aa SGNH1 from Aphanizomenon flos-aquae expressed as insoluble inclusion bodies, refAbstract=null), Reference(id=1226592773399294847, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2019, volume=41, issue=4/5, pageStart=591, pageEnd=604, url=null, language=null, rfNumber=[17], rfOrder=16, authorNames=HONG LG, JIAN SL, HUO YY, CHENG H, HU XJ, LI JX, CUI HL, XU XW, journalName=Biotechnology Letters, refType=null, unstructuredReference=HONG LG, JIAN SL, HUO YY, CHENG H, HU XJ, LI JX, CUI HL, XU XW. A novel SGNH family hydrolase Ali5 with thioesterase activity and a GNSL motif but without a classic GDSL motif from Altererythrobacter ishigakiensis [J]. Biotechnology Letters, 2019, 41(4/5): 591-604., articleTitle=A novel SGNH family hydrolase Ali5 with thioesterase activity and a GNSL motif but without a classic GDSL motif from Altererythrobacter ishigakiensis, refAbstract=null), Reference(id=1226592773504152450, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2022, volume=13, issue=null, pageStart=922506, pageEnd=null, url=null, language=null, rfNumber=[18], rfOrder=17, authorNames=CHEN CH, YU G, GUO ZY, YANG QH, SU WF, XIE QF, YANG GD, REN YF, LI H, journalName=Frontiers in Microbiology, refType=null, unstructuredReference=CHEN CH, YU G, GUO ZY, YANG QH, SU WF, XIE QF, YANG GD, REN YF, LI H. Expression, characterization, fermentation, immobilization, and application of a novel esterase Est804 from metagenomic library in pesticide degradation[J]. Frontiers in Microbiology, 2022, 13: 922506., articleTitle=Expression, characterization, fermentation, immobilization, and application of a novel esterase Est804 from metagenomic library in pesticide degradation, refAbstract=null), Reference(id=1226592773663536006, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2021, volume=31, issue=3, pageStart=483, pageEnd=491, url=null, language=null, rfNumber=[19], rfOrder=18, authorNames=JO E, KIM J, LEE A, MOON K, CHA J, journalName=Journal of Microbiology and Biotechnology, refType=null, unstructuredReference=JO E, KIM J, LEE A, MOON K, CHA J. Identification and characterization of a novel thermostable GDSL-type lipase from Geobacillus thermocatenulatus [J]. Journal of Microbiology and Biotechnology, 2021, 31(3): 483-491., articleTitle=Identification and characterization of a novel thermostable GDSL-type lipase from Geobacillus thermocatenulatus, refAbstract=null), Reference(id=1226592773772587917, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2021, volume=179, issue=null, pageStart=576, pageEnd=585, url=null, language=null, rfNumber=[20], rfOrder=19, authorNames=GULERIA S, JAIN R, SINGH D, KUMAR S, journalName=International Journal of Biological Macromolecules, refType=null, unstructuredReference=GULERIA S, JAIN R, SINGH D, KUMAR S. A thermostable Fe/Mn SOD of Geobacillus sp. PCH100 isolated from glacial soil of Indian trans-Himalaya exhibits activity in the presence of common inhibitors[J]. International Journal of Biological Macromolecules, 2021, 179: 576-585., articleTitle=A thermostable Fe/Mn SOD of Geobacillus sp. PCH100 isolated from glacial soil of Indian trans-Himalaya exhibits activity in the presence of common inhibitors, refAbstract=null), Reference(id=1226592773877445521, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2020, volume=599, issue=null, pageStart=113741, pageEnd=null, url=null, language=null, rfNumber=[21], rfOrder=20, authorNames=AKRAM F, HAQ IU, journalName=Analytical Biochemistry, refType=null, unstructuredReference=AKRAM F, HAQ IU. Overexpression and characterization of TnCel12B, a hyperthermophilic GH12 endo-1,4-β-glucanase cloned from Thermotoga naphthophila RKU-10T [J]. Analytical Biochemistry, 2020, 599: 113741., articleTitle=Overexpression and characterization of TnCel12B, a hyperthermophilic GH12 endo-1,4-β-glucanase cloned from Thermotoga naphthophila RKU-10T, refAbstract=null), Reference(id=1226592773994886039, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2016, volume=63, issue=1, pageStart=41, pageEnd=50, url=null, language=null, rfNumber=[22], rfOrder=21, authorNames=YUAN DJ, LAN DM, XIN RP, YANG B, WANG YH, journalName=Biotechnology and Applied Biochemistry, refType=null, unstructuredReference=YUAN DJ, LAN DM, XIN RP, YANG B, WANG YH. Screening and characterization of a thermostable lipase from marine Streptomyces sp. strain W007[J]. Biotechnology and Applied Biochemistry, 2016, 63(1): 41-50., articleTitle=Screening and characterization of a thermostable lipase from marine Streptomyces sp. strain W007, refAbstract=null), Reference(id=1226592774103937944, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2024, volume=26, issue=3, pageStart=447, pageEnd=459, url=null, language=null, rfNumber=[23], rfOrder=22, authorNames=RODRÍGUEZ-MEJÍA JL, HIDALGO-MANZANO IA, MURIEL-MILLÁN LF, RIVERA-GOMEZ N, SAHONERO-CANAVESI DX, CASTILLO E, PARDO-LÓPEZ L, journalName=Marine Biotechnology, refType=null, unstructuredReference=RODRÍGUEZ-MEJÍA JL, HIDALGO-MANZANO IA, MURIEL-MILLÁN LF, RIVERA-GOMEZ N, SAHONERO-CANAVESI DX, CASTILLO E, PARDO-LÓPEZ L. A novel thermo-alkaline stable GDSL/SGNH esterase with broad substrate specificity from a deep-sea Pseudomonas sp.[J]. Marine Biotechnology, 2024, 26(3): 447-459., articleTitle=A novel thermo-alkaline stable GDSL/SGNH esterase with broad substrate specificity from a deep-sea Pseudomonas sp, refAbstract=null), Reference(id=1226592774204601242, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, doi=null, pmid=null, pmcid=null, year=2022, volume=69, issue=5, pageStart=2176, pageEnd=2194, url=null, language=null, rfNumber=[24], rfOrder=23, authorNames=RAFEEQ H, HUSSAIN A, SHABBIR S, ALI S, BILAL M, SHER F, IQBAL HMN, journalName=Biotechnology and Applied Biochemistry, refType=null, unstructuredReference=RAFEEQ H, HUSSAIN A, SHABBIR S, ALI S, BILAL M, SHER F, IQBAL HMN. Esterases as emerging biocatalysts: mechanistic insights, genomic and metagenomic, immobilization, and biotechnological applications[J]. 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A: Phylogenetic analysis of EstE (◆) (The tree was constructed using the neighbor-joining method in MEGA 7.0, and all sequences were retrieved from the NCBI database; Numbers on branches indicate bootstrap support values from 1 000 replicates); B: Sequence alignment analysis of EstE with related SGNH family hydrolases using ClustalW and ESPript (Conserved sequence blocks (I, II, III, and V) and characteristic residues are indicated by black boxes and triangles, respectively; The catalytic triad and oxyanion hole are highlighted with red and green triangles, respectively)., figureFileSmall=pxk1MwT8Cg79pUkEnwmFpA==, figureFileBig=LKgIeBA+oqQE7ANC+JXvLQ==, tableContent=null), ArticleFig(id=1226592761768489571, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=CN, label=图1, caption=EstE的系统发育及序列分析。A：EstE (◆)系统发育分析(使用MEGA 7.0邻接法模型建立系统发育树，所有序列均从NCBI数据库检索得到，分支上的数字表示1 000次自举法检验的可靠性百分比)；B：使用ClustalW和ESPript对EstE及相关SGNH家族水解酶的序列比对分析[黑色方框表示4个保守序列区块(I、II、III和V)，黑色三角形表示每个保守区块中的特征氨基酸，催化三联体和氧阴离子空穴分别用红色和绿色三角形表示]。, figureFileSmall=pxk1MwT8Cg79pUkEnwmFpA==, figureFileBig=LKgIeBA+oqQE7ANC+JXvLQ==, tableContent=null), ArticleFig(id=1226592761919484525, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=EN, label=Figure 2, caption=The 3D structure and surface electrostatic potential analysis of EstE. A: The 3D structure of EstE predicted by AlphaFold (Alpha helices, beta sheets, and random coils are colored green, red, and yellow, respectively; The catalytic triad (Ser16, Asp194, His224) is shown in magenta); B: Surface electrostatic potential of EstE analyzed by PyMOL (Blue and red regions represent positive and negative charges, respectively)., figureFileSmall=yd2n5oQfi71nFgXDMx3Dvw==, figureFileBig=nwTJXxrcbeLtEkNlPT7nHg==, tableContent=null), ArticleFig(id=1226592762024342133, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=CN, label=图2, caption=EstE三维结构和表面静电势分析。A：AlphaFold预测的EstE三维结构模型[绿色表示α-螺旋，红色表示β-折叠，黄色表示无规卷曲，洋红色表示催化三联体(Ser16、Asp194、His224)]；B：通过PyMOL分析的EstE表面静电势(蓝色和红色区域分别代表正电荷和负电荷分布)。, figureFileSmall=yd2n5oQfi71nFgXDMx3Dvw==, figureFileBig=nwTJXxrcbeLtEkNlPT7nHg==, tableContent=null), ArticleFig(id=1226592762179531386, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=EN, label=Figure 3, caption=SDS-PAGE of EstE. Lane M: Standard marker proteins; Lane 1: Cell lysate of EstE; Lane 2: Supernatant of cell lysate of EstE; Lane 3: Purified EstE., figureFileSmall=+klNEMWqiVvY0Hs+s2dj+w==, figureFileBig=IuYwBb4OflHK0Y83uzP3UQ==, tableContent=null), ArticleFig(id=1226592762296971904, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=CN, label=图3, caption=酯酶EstE的SDS-PAGE分析。泳道M：标准标记蛋白；泳道1：EstE细胞裂解液；泳道2：EstE细胞裂解液的上清液；泳道3：纯化的EstE。, figureFileSmall=+klNEMWqiVvY0Hs+s2dj+w==, figureFileBig=IuYwBb4OflHK0Y83uzP3UQ==, tableContent=null), ArticleFig(id=1226592762414412427, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=EN, label=Figure 4, caption=Substrate specificity and specific enzyme activity of EstE. A: Specific enzyme activity of EstE; B:Substrate specificity of EstE. Different letters indicate significant differences (P<0.05)., figureFileSmall=+FGNo+RWLwiglAKJRDZXaA==, figureFileBig=uegPjyyzBapduggECQLiqw==, tableContent=null), ArticleFig(id=1226592763794338451, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=CN, label=图4, caption=EstE的底物特异性及比酶活。A：EstE的比酶活；B：EstE的底物特异性。不同字母表示显著性差异(P<0.05)。, figureFileSmall=+FGNo+RWLwiglAKJRDZXaA==, figureFileBig=uegPjyyzBapduggECQLiqw==, tableContent=null), ArticleFig(id=1226592765757272726, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=EN, label=Figure 5, caption=Effects of different pHs on enzyme activity of EstE. A: The optimal pH of esterase EstE (The activity was determined in 50 mmol/L buffers: sodium citrate buffer (pH 5.5-7.0, ■), Tris-HCl buffer (pH 7.0-9.0, ●), and KH2PO4-KOH buffer (pH 9.0-10.5, ▲)); B: The pH stability of EstE (The enzyme was incubated in Tris-HCl buffer (pH 8.5) for 100 h, and the activity of untreated EstE was defined as 100%)., figureFileSmall=pGg4e7PYLol3oie1FRaj3A==, figureFileBig=lLzU8+yGyb0lC1JP7VYc8g==, tableContent=null), ArticleFig(id=1226592766021513884, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=CN, label=图5, caption=不同pH对酯酶EstE活性的影响。A：EstE的最适pH值[在50 mmol/L不同缓冲液中测定酯酶活性：柠檬酸钠缓冲液(pH 5.5-7.0，■)，Tris-HCl缓冲液(pH 7.0-9.0，●)和KH2PO4-KOH缓冲液(pH 9.0-10.5，▲)]；B：EstE的pH稳定性[EstE在Tris-HCl缓冲液(pH 8.5)中孵育100 h，未经处理的EstE酶活性定义为100%]。, figureFileSmall=pGg4e7PYLol3oie1FRaj3A==, figureFileBig=lLzU8+yGyb0lC1JP7VYc8g==, tableContent=null), ArticleFig(id=1226592766151537312, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=EN, label=Figure 6, caption=Effects of different temperatures on enzyme activity of EstE. A: The optimal temperature of EstE (The activity was determined in Tris-HCl buffer (pH 8.5) at various temperatures (15-65 ℃) and the error bar represents the standard deviation (n=3)); B: Thermostability of EstE at 40 ℃ for 26 h; C: Thermostability of EstE at 100 ℃ for 10 h., figureFileSmall=eFTyEDYMfLRibY3QP5bKQA==, figureFileBig=ADksGkCHezHQ1I5/TBZhmQ==, tableContent=null), ArticleFig(id=1226592766302532262, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=CN, label=图6, caption=不同温度对酯酶EstE活性的影响。A：EstE的最适温度[在Tris-HCl缓冲液(pH 8.5)中，以不同温度(15-65 ℃)测定酶活性，误差条表示标准差(n=3)]；B：EstE在40 ℃下孵育26 h的热稳定性；C：EstE在100 ℃下孵育10 h的热稳定性。, figureFileSmall=eFTyEDYMfLRibY3QP5bKQA==, figureFileBig=ADksGkCHezHQ1I5/TBZhmQ==, tableContent=null), ArticleFig(id=1226592766461915822, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=EN, label=Figure 7, caption=Thermal inactivation kinetics of EstE at 40 ℃ and 100 ℃. A: Thermal inactivation kinetics at 40 ℃; B: Thermal inactivation kinetics at 100 ℃. The linear relationship between time and the logarithm of relative enzyme activity of EstE (ln E/E0, where E0 and E represent the initial enzyme activity without heat treatment and the residual enzyme activity after heat treatment for different times at 40 or 100 ℃, respectively)., figureFileSmall=jZ0HOUFZhr1cx9hCg6JxAg==, figureFileBig=YjcYp7rhSRPZhtRMQMZ6cg==, tableContent=null), ArticleFig(id=1226592766642270900, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=CN, label=图7, caption=40 ℃和100 ℃条件下EstE的热失活动力学。A：40 ℃下EstE的热失活动力学；B：100 ℃下EstE的热失活动力学。EstE相对酶活性的对数与时间的线性关系(ln E/E0，其中E0和E分别表示未经热处理的初始酶活性和在40 ℃或100 ℃下不同时间热处理后的剩余酶活性)。, figureFileSmall=jZ0HOUFZhr1cx9hCg6JxAg==, figureFileBig=YjcYp7rhSRPZhtRMQMZ6cg==, tableContent=null), ArticleFig(id=1226592766789071548, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=EN, label=Figure 8, caption=Effects of metal ions, detergents and organic solvents on enzyme activity of EstE. A: Influences of different metal ions (1 mmol/L and 10 mmol/L) on EstE; B: Influences of different detergents (0.1% and 1%) on EstE; C: Influences of various organic solvents (15% and 30%) on EstE. Letters indicate statistical significance among groups (P<0.05)., figureFileSmall=rDy4JIXdMEZG8iKcC387GA==, figureFileBig=1Ju/4A5CxrkeXIoN0f1biw==, tableContent=null), ArticleFig(id=1226592766898123456, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=CN, label=图8, caption=金属离子、去垢剂和有机溶剂对EstE酶活性的影响。A：不同金属离子(1 mmol/L和10 mmol/L)对EstE活性的影响；B：不同去垢剂(0.1%和1%)对EstE活性的影响；C：不同有机溶剂(15%和30%)对EstE活性的影响。字母表示组间差异具有统计学意义(P<0.05)。, figureFileSmall=rDy4JIXdMEZG8iKcC387GA==, figureFileBig=1Ju/4A5CxrkeXIoN0f1biw==, tableContent=null), ArticleFig(id=1226592766990398149, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=EN, label=Table 1, caption=

The comparison of nucleotide composition of estE′ and estE

, figureFileSmall=null, figureFileBig=null, tableContent=

核苷酸

Nucleotide

优化前(estE)

Before optimization

优化后(estE′)

After optimization

计数

Count

占比

Proportion

(%)

计数

Count

占比

Proportion(%)

腺嘌呤A10712.312214.0
胞嘧啶C31936.722726.1
鸟嘌呤G32337.133238.2
胸腺嘧啶T12113.918921.7
C+G64273.855964.3
A+T22826.231135.7
), ArticleFig(id=1226592768370324171, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=CN, label=表1, caption=

酯酶基因 estE 人工优化前后的核苷酸序列比较表

, figureFileSmall=null, figureFileBig=null, tableContent=

核苷酸

Nucleotide

优化前(estE)

Before optimization

优化后(estE′)

After optimization

计数

Count

占比

Proportion

(%)

计数

Count

占比

Proportion(%)

腺嘌呤A10712.312214.0
胞嘧啶C31936.722726.1
鸟嘌呤G32337.133238.2
胸腺嘧啶T12113.918921.7
C+G64273.855964.3
A+T22826.231135.7
), ArticleFig(id=1226592768559067858, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=EN, label=Table 2, caption=

Comparison of enzyme properties of EstE with other enzymes from the GDS(L) family

, figureFileSmall=null, figureFileBig=null, tableContent=

酯酶

Enzyme

来源

Source

最适温度

Optimal temperature

(℃)

最适pH

Optimal pH

最适底物

Optimal substrate

比酶活

Specific enzyme activity (U/mg)

参考文献

References

AaSGNH1Aphanizomenon flos-aquae108.0-9.0C21.70[16]
Ali5Altererythrobacter ishigakiensis407.5C414.60[17]
Est804Metagenomic library in pesticide degradation378.0C848.97[18]
Lip29Geobacillus thermocatenulatus509.5C142.27[19]
Est29Geobacillus thermocatenulatus556.0C80.92[19]
EstEStreptomyces griseus408.5C261.03This study
), ArticleFig(id=1226592768689091288, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236833328181880, language=CN, label=表2, caption=

EstE与其他GDS(L)家族酯酶的酶学性质比较

, figureFileSmall=null, figureFileBig=null, tableContent=

酯酶

Enzyme

来源

Source

最适温度

Optimal temperature

(℃)

最适pH

Optimal pH

最适底物

Optimal substrate

比酶活

Specific enzyme activity (U/mg)

参考文献

References

AaSGNH1Aphanizomenon flos-aquae108.0-9.0C21.70[16]
Ali5Altererythrobacter ishigakiensis407.5C414.60[17]
Est804Metagenomic library in pesticide degradation378.0C848.97[18]
Lip29Geobacillus thermocatenulatus509.5C142.27[19]
Est29Geobacillus thermocatenulatus556.0C80.92[19]
EstEStreptomyces griseus408.5C261.03This study
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灰色链霉菌热碱稳定性酯酶的异源表达和酶学性质
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李全发 1 , 房金鑫 1 , 喻娇 1 , 陈菁菁 1 , 王宝娟 1, 2
微生物学报 | 研究报告 2025,65(5): 2201-2213
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微生物学报 | 研究报告 2025, 65(5): 2201-2213
灰色链霉菌热碱稳定性酯酶的异源表达和酶学性质
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李全发1, 房金鑫1, 喻娇1, 陈菁菁1, 王宝娟1, 2
作者信息
  • 1.安徽师范大学 生命科学学院，安徽 芜湖
  • 2.分子酶学与重大代谢性疾病机理研究安徽省重点实验室，安徽 芜湖
Heterologous expression and enzymatic characterization of a thermostable and alkaline-stable esterase from Streptomyces griseus
Quanfa LI1, Jinxin FANG1, Jiao YU1, Jingjing CHEN1, Baojuan WANG1, 2
Affiliations
  • 1.College of Life Sciences, Anhui Normal University, Wuhu, Anhui, China
  • 2.Anhui Provincial Key Laboratory of Molecular Enzymology and Mechanism of Major Metabolic Diseases, Wuhu, Anhui, China
出版时间: 2025-05-04 doi: 10.13343/j.cnki.wsxb.20240718
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摘要
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【目的】通过在大肠杆菌中异源表达，获得一种来源于灰色链霉菌(Streptomyces griseus)的新型酯酶EstE，并系统评估其热稳定性、碱稳定性以及不同添加剂(金属离子、去垢剂和有机溶剂)对酶活性的影响，以探索其在工业应用中的潜力。【方法】对灰色链霉菌中酯酶基因estE进行人工优化，合成编码相同氨基酸序列的酯酶基因estE'，将其与载体pET-28b(+)连接后，构建含该酯酶基因的重组质粒。通过IPTG诱导表达，采用Co2+亲和层析纯化得到酯酶EstE，随后利用对硝基苯酚法对该酶的酶学性质进行鉴定，并对其进行生物信息学分析。【结果】EstE由289个氨基酸组成，分子量约为31.6 kDa，属于GDS(L)家族，其催化三联体由Ser16、Asp194、His224构成。该酶的最适温度和最适pH分别为40 ℃和pH 8.5，以对硝基苯酚乙酸酯为底物时催化活性最高，比酶活为61.03 U/mg。在热稳定性方面，酯酶EstE在40 ℃和100 ℃条件下的半衰期分别为156.11 h和2.67 h；在碱稳定性方面，其在pH 8.5下孵育100 h后，仍能维持80%以上的相对酶活性。在有机溶剂耐受性方面，当存在30% DMSO时，其酶活基本不变。【结论】通过异源表达，成功获得了一种来源于灰色链霉菌的新型酯酶EstE。该酶具有优异的催化特性、热稳定性、碱稳定性和有机溶剂耐受性，为其在工业领域中的应用奠定了基础。

水解酶  /  异源表达  /  热稳定性  /  碱稳定性

[Objective] This study characterized a novel esterase EstE from Streptomyces griseus by heterologous expression in Escherichia coli and systematically evaluates its thermostability, alkaline stability, and the effects of various additives (metal ions, detergents, and organic solvents) on its enzymatic activity to explore its potential for industrial applications. [Methods] We synthesized the gene estE' encoding the same amino acid sequence as the native gene by optimizing the original sequence of estE from S. griseus. We then constructed the recombinant plasmid carrying the optimized gene by ligating the gene into the pET-28b(+) vector. The esterase EstE was then expressed under the induction of IPTG and purified via Co2+ affinity chromatography. Furthermore, the enzymatic properties of the purified EstE were determined by the p-nitrophenol method, and bioinformatics analysis was performed for this enzyme. [Results] EstE consisted of 289 amino acid residues, with a molecular weight of 31.6 kDa. It belonged to the GDS(L) family, with Ser16, Asp194, and His224 forming its catalytic triad. The enzyme showed the optimal activity at 40 ℃ and pH 8.5, with the highest catalytic efficiency (specific activity of 61.03 U/mg) observed in the case of p-nitrophenyl acetate as a substrate. EstE demonstrated robust thermostability, with the relative activity of 50% after 156.11 h of incubation at 40 ℃ and 2.67 h of incubation at 100 ℃. Moreover, it showed excellent alkaline stability, with the relative activity exceeding 80% after incubation at pH 8.5 for 100 h. In addition, this enzyme exhibited excellent tolerance to organic solvents, maintaining stable activity in the presence of 30% DMSO. [Conclusion] A novel esterase EstE from S. griseus is successfully obtained through heterologous expression, demonstrating excellent catalytic properties, thermostability, alkaline stability, and organic solvent tolerance, positioning it as a promising candidate for industrial applications.

hydrolase  /  heterologous expression  /  thermostability  /  alkaline stability
李全发, 房金鑫, 喻娇, 陈菁菁, 王宝娟. 灰色链霉菌热碱稳定性酯酶的异源表达和酶学性质. 微生物学报, 2025 , 65 (5) : 2201 -2213 . DOI: 10.13343/j.cnki.wsxb.20240718
Quanfa LI, Jinxin FANG, Jiao YU, Jingjing CHEN, Baojuan WANG. Heterologous expression and enzymatic characterization of a thermostable and alkaline-stable esterase from Streptomyces griseus[J]. Acta Microbiologica Sinica, 2025 , 65 (5) : 2201 -2213 . DOI: 10.13343/j.cnki.wsxb.20240718
正文
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酯酶(EC 3.1.1.X, esterase)广泛分布于动物、植物和微生物中，在水相中能够催化酯键的水解反应，将脂类分解成脂肪酸和醇；在非水相中则催化酯化或转酯反应生成酯键[1-2]。酯酶在结构上隶属于α/β水解酶超家族，具有由Ser-His-Asp构成的催化三联体，其中亲核氨基酸Ser残基通过氢键与His相连。由于酯酶在催化底物水解和合成时不需要辅助因子，并且在催化反应中具有化学选择性、立体特异性和区域专一性等优点，使其被广泛应用于精细化工、药物合成、生物修复等领域[3-4]。基于酯酶氨基酸序列同源性和部分生化特征，1999年Arpigny和Jaeger[5]首次将细菌酯酶分为8大家族：true lipase (Family I)、GDS(L) Family (Family II)、Family III、Hormone-sensitive lipase (HSL Family/Family IV)、Family V-VIII。近年来，随着基因组测序和蛋白质功能的深入研究，微生物酯酶已被扩展至Family XXI[2,6-7]，其中，GDS(L) Family酯酶并不表现出传统的GXSXG基序，而是显示出包含活性位点丝氨酸残基的GDSL基序。此外，GDS(L) Family的一个亚群被划分为SGNH水解酶，该亚群成员在4个保守块I、II、III和V中分别具有保守的Ser、Gly、Asn和His残基。值得一提的是，II区中的保守Gly与III区中的保守Asn协同作用，作为氧阴离子空穴的质子供体，参与催化过程中的质子转移；而位于V区的碱性His则通过去质子化作用，增强了活性位点丝氨酸的亲核性，从而促进了酶催化反应的进行，这一系列精细的分子机制共同构成了GDS(L)家族中SGNH水解酶亚群独特的催化特性[7]。
酶作为一种生物催化剂，具有催化效率高、环境友好等优点。工业生产常面临严苛的反应条件，如高温、强碱和有机溶剂等恶劣环境，这些条件往往会严重损害酶的催化活性，从而极大地限制了酶在工业中的应用[8-9]。因此，开发具有优良热稳定性、碱稳定性和有机溶剂耐受性的酯酶具有重要的理论和现实意义。尽管酯酶存在于几乎所有生物中，但与动植物酯酶相比，来自微生物的酯酶具有催化活性多样、产量高、遗传操作简单、供应稳定以及生长速度快等优点，因此微生物酯酶是工业应用酯酶的主要来源[4]。链霉菌是一类存在于土壤中的革兰氏阳性菌，可产生包括酯酶在内的大量具生物活性的次级代谢产物。然而，截至目前，只有极少数的链霉菌酯酶被分离鉴定，且表现出独特的酶学性质，如较高的有机溶剂耐受性、耐碱性、耐高温或嗜冷性等[10-11]。因此，链霉菌可以作为开发具有优良酶学特性的新型酯酶的理想来源。本研究以灰色链霉菌(Streptomyces griseus)酯酶EstE为研究对象，人工合成酯酶基因estE′，构建含该基因的重组质粒并成功实现其在大肠杆菌中的异源表达。随后采用MEGA和AlphaFold对酯酶EstE进行系统发育分析和结构预测；进一步利用对硝基苯酚法对酯酶EstE的底物特异性、最适pH、最适温度、热稳定性、碱稳定性等酶学性质进行系统鉴定，旨在挖掘一种具有优良酶学性能的新型链霉菌酯酶。
1 材料与方法
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1.1 菌株和质粒
克隆宿主菌大肠杆菌(Escherichia coli) DH5α及含有氯霉素(chloramphenicol, Cam)抗性的表达宿主菌E. coli Rosetta(DE3)均购自生工生物工程(上海)股份有限公司。目的基因estE为灰色链霉菌染色体基因座SGR_RS31360所对应的酯酶基因，优化后的目的基因estE′由南京金斯瑞生物科技有限公司人工合成，同时利用携带卡那霉素(kanamycin, Kan)抗性的质粒pET-28b(+)构建重组表达质粒pET-28b(+)-estE′用于酯酶的异源表达。
1.2 主要试剂
对硝基苯酚(p-nitrophenol, pNP)及不同链长的对硝基苯酚酯类[对硝基苯酚乙酸酯(p-nitrophenyl acetate, pNPA2)、对硝基苯酚丁酸酯(p-nitrophenyl butyrate, pNPB4)、对硝基苯酚己酸酯(p-nitrophenyl caproate, pNPC6)、对硝基苯酚辛酸酯(p-nitrophenyl caprylate, pNPC8)、对硝基苯酚奎酸酯(p-nitrophenyl caprate, pNPC10)、对硝基苯酚月桂酸酯(p-nitrophenyl laurate, pNPL12)、对硝基苯酚豆蔻酸酯(p-nitrophenyl myristate, pNPM14)、对硝基苯酚棕榈酸酯(p-nitrophenyl palmitate, pNPP16)]均购自Sigma-Aldrich公司；质粒提取试剂盒、胶回收试剂盒均购自Axygen公司；Co2+亲和层析树脂购自TaKaRa公司；层析柱、异丙基硫代半乳糖苷(IPTG)、Brandford试剂、三羟甲基氨基甲烷(Tris)以及其他试剂均购自生工生物工程(上海)股份有限公司。
1.3 生物信息学分析
从NCBI (http://www.ncbi.nlm.nih.gov/)下载EstE氨基酸序列(WP_003970591)，利用BLASTp进行EstE氨基酸序列相似性检索并下载相似性序列，运用ClustalW和ESPript 3.0 (http://espript.ibcp.fr/ESPript/cgi-bin/ESPript.cgi)进行氨基酸多重序列比对分析；运用MEGA 7.0邻接法将从NCBI下载的来源于其他8个酯酶家族的氨基酸序列以及EstE氨基酸序列一起构建系统发育树。利用AlphaFold (https://www.alphafold.ebi.ac.uk/)对EstE进行三维结构建模，使用PyMOL对生成的三维结构模型进行分析。
1.4 重组质粒pET-28b(+)-estE′ 的构建
将人工设计合成的酯酶基因estE′和表达载体pET-28b(+)分别用Nde I和Xho I双酶切，酶切条件为37 ℃、6 h。酶切后的产物通过琼脂糖凝胶电泳分离，并进行胶回收。随后将纯化后的目标基因estE′与载体pET-28b(+)的酶切产物混合，16 ℃连接过夜，构建重组质粒pET-28b(+)-estE′。将连接产物转化入大肠杆菌感受态细胞E. coli DH5α，在含有Kan抗性的LB平板上37 ℃倒置培养过夜。挑选转化后的阳性单克隆菌株在37 ℃下进行摇瓶培养，并提取质粒，通过琼脂糖凝胶电泳鉴定，随后送生工生物工程(上海)股份有限公司进一步测序验证。
1.5 重组酯酶EstE的异源表达及分离纯化
将测序验证正确的重组质粒pET-28b(+)-estE'转化入表达菌株E. coli Rosetta(DE3)，构建重组工程菌Rosetta(DE3) pLysS/pET28b-estE′，并通过含Kan和Cam双抗的LB固体培养基筛选出抗性重组工程菌。随后挑取单菌落至装有5 mL双抗的LB液体培养基中，37 ℃、225 r/min培养过夜。吸取1 mL菌液至含双抗的100 mL LB液体培养基中，37 ℃、225 r/min摇床培养至OD600达到0.4-0.6。向培养物中加入诱导剂IPTG至终浓度为0.1 mmol/L，在20 ℃、180 r/min摇床中低温诱导表达20-24 h。随后4 ℃、5 000 r/min离心5 min收集菌体，超声波破碎30 min (超声破碎仪参数：工作时间1 s，间隔时间2 s，功率87 W)。破碎后菌液4 ℃、5 000 r/min离心5 min，上清液加入含Co2+树脂的亲和层析柱中充分混匀，30 min后用缓冲液冲洗去除杂蛋白，最后用洗脱缓冲液洗脱与层析柱结合能力强的目的蛋白，得到羧酸酯酶EstE。利用12%的聚丙烯酰胺凝胶电泳(SDS-PAGE)检测酯酶的纯化效果。蛋白质浓度的测定采用Bradford法，以牛血清白蛋白为标准品。
1.6 重组酯酶EstE的酶学性质鉴定
酯酶EstE可以催化底物对硝基苯酚酯类水解，并在410 nm处生成有特异性吸收峰的产物对硝基苯酚(pNP)。利用紫外-可见光分光光度计监测吸光值在对应波长处的变化，并通过测定反应体系中pNP的释放量，确定重组酯酶EstE的酶活性。将Tris-HCl缓冲液(pH 8.5，980 μL)、pNPA2 (10 μL)和纯化酯酶EstE (10 μL)混合均匀后，置于25 ℃水浴锅中温育5 min，立即测定OD410。以未添加酶的反应物为对照，所有平行实验均独立重复3次。
1.6.1 底物特异性
在25 ℃、pH 8.5条件下，检测EstE对不同碳链长的对硝基苯酚酯类(C2-C16)的水解活性，并根据对硝基苯酚标准曲线计算酶活。所测得的酶活最大值所对应的对硝基苯酚酯类为该酶的最适底物。一个酶活单位(U)定义为在pH 8.5和25 ℃条件下，单位时间内每产生1 μmol对硝基苯酚所需要的酶量。
1.6.2 pH对酶活性的影响
在25 ℃条件下，以pNPA2为底物测定在不同pH条件下EstE的酶活性。缓冲液包括50 mmol/L柠檬酸缓冲液(pH 5.5-7.0)、50 mmol/L Tris-HCl缓冲液(pH 7.0-9.0)和50 mmol/L K2HPO4-KOH缓冲液(pH 9.0-10.5)。以所测得的最适pH条件下的酶活力为100%，计算其他pH缓冲液中的相对酶活。为检测酯酶EstE的碱稳定性，将EstE置于4 ℃、pH 8.5条件下孵育一段时间后，以pNPA2为底物，并在pH 8.5和25 ℃条件下，检测其残余酶活力，以未经碱处理的酶活力定义为100%，计算不同碱处理时间下的相对酶活。
1.6.3 温度对酶活性的影响
在pH 8.5条件下，以pNPA2为底物，设置温度梯度为10-65 ℃，采用标准反应体系测定酯酶EstE的最适反应温度，以测定的最适温度下的酶活力为100%，计算其他温度下的相对酶活。为进一步检测酯酶EstE的热稳定性，酶分别在40 ℃和100 ℃孵育一段时间后，以pNPA2为底物，并在pH 8.5和25 ℃条件下，采用标准检测体系检测其残余酶活力，将未经热处理的酶活力定义为100%。随后以剩余酶活性的对数ln E/E0与时间(t)作图，计算EstE的热失活动力学参数[12]。
1.6.4 不同添加剂对酶活性的影响
为检测金属离子和化学试剂对EstE活性的影响，向标准反应体系中分别添加一定浓度的金属离子(KCl、NaCl、LiCl、CoCl2、ZnSO4、CaCl2、NiSO4、BaCl2、MnCl2、CuCl2、MgCl2、HgCl2、AlCl3)、有机溶剂[N,N-二甲基甲酰胺(N,N-dimethylformamide, DMF)、二甲基亚砜(dimethyl sulfoxide, DMSO)、甲醇、乙醇、乙腈、丙酮]、去垢剂[十六烷基三甲基溴化铵(cetyltrimethylammonium bromide, CTAB)、十二烷基磺酸钠(sodium dodecyl sulfate, SDS)、吐温-20 (Tween-20)、吐温-80 (Tween-80)、聚乙二醇辛基苯基醚(triton X-100)]，在25 ℃、pH 8.5条件下，以pNPA2为底物反应5 min后检测EstE残余酶活，以不添加任何化学试剂的EstE活性为100%。其中，金属离子的检测浓度为1 mmol/L和10 mmol/L，有机溶剂的检测体积分数分别为15%和30%，去垢剂的检测体积分数分别为0.1%和1%。
2 结果与分析
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2.1 酯酶基因 estE 的优化及合成
将灰色链霉菌染色体(NC_010572)基因座SGR_RS31360所编码的酯酶基因命名为estE。该基因在染色体上核苷酸序列起始位点为7 453 938，终止位点为7 454 807，全长870 bp，所编码蛋白质全长289个氨基酸，预测其分子量为31.6 kDa。在GenBank中蛋白质的编号为WP_003970591，将其命名为酯酶EstE。由于该基因G+C含量高，占比为73.8%，这使其在大肠杆菌中异源表达受到限制。因此，本研究以大肠杆菌偏爱密码子代替链霉菌密码子和降低G+C含量两方面对酶基因estE进行优化。优化后酯酶基因命名为estE'，其基因长度和所编码的氨基酸序列保持不变，但与原始序列相比，G+C含量从73.8%降至64.3% (表1)。
2.2 酯酶EstE的生物信息学分析
为鉴定EstE在分子水平上的家族亲缘关系，对已建立的8个细菌酯酶家族的22个代表性成员与EstE的氨基酸序列一起进行发育树分析。结果显示EstE在分子发育树上属于酯酶Family II (图1A)。进一步对该家族的酯酶进行多重序列比对(图1B)，EstE氨基酸序列中含有SGNH家族水解酶特异性4个保守片段(I、II、III和V)，其中包含特征性残基(Ser、Gly、Asn和His) 4个。活性位点Ser16位于区域I的GDS基序中，DXXH基序位于区域V中。区域II和III中的保守残基(Gly、Asn)参与了氧阴离子空穴的形成[13]。进一步基于AlphaFold构建EstE的三维结构(图2A)，EstE是一种球形蛋白，由11个α-螺旋紧密围绕由7条β-链组成的β-片层形成。Ser16位于α1螺旋上，Asp194位于α9和α10中间，His224位于α10螺旋上，它们在空间上聚集形成位于三维结构表面凹槽中的催化三联体。此外，对EstE的静电势进行分析(图2B)，代表正电荷的蓝色区域和负电荷的红色区域几乎相等地覆盖在蛋白质表面，表明其表面正负电荷分布较为均匀[14]。
2.3 酯酶EstE的表达及纯化
重组大肠杆菌工程菌Rosetta(DE3) pLysS/pET28b-estE'低温诱导表达20-24 h后，通过离心收集菌体并进行破碎。得到的EstE粗酶液经Co2+亲和层析柱纯化后，纯化产物进行12% SDS-PAGE分析。结果显示在靠近35 kDa处有一条明显的主条带(图3)。酯酶EstE的分子量理论计算值为31.6 kDa，由于重组质粒上含有6个His标签，含该标签的EstE分子量约为32.5 kDa，与蛋白质电泳条带基本一致。由此可见，酯酶EstE成功在大肠杆菌中实现了异源表达，并获得了高纯度的蛋白产物。
2.4 酯酶EstE的最适底物
以不同碳链长度(C2-C16)的对硝基苯酚酯为底物，检测酯酶EstE的酶活性并计算比活力。结果如图4所示，EstE偏好水解短链酯类(C2-C4)，其最适反应底物为pNPA2 (C2)，且随着碳链长度的增加，EstE的相对酶活力逐渐降低。当底物碳链长度大于6 (C6-C16)时，EstE的相对酶活力均低于10% (图4A)；以pNPA2和pNPB4为底物时，EstE的比酶活分别为61.03 U/mg和32.46 U/mg (图4B)。
2.5 酯酶EstE的最适pH和碱稳定性
在25 ℃条件下，以不同pH值的缓冲液(pH 5.5-10.5)测定EstE的酶活性。结果显示，EstE的最适pH为8.5，在pH 7.5-10.5范围内，酶活力能保持在60%以上(图5A)。进一步对EstE的碱稳定性进行鉴定，结果如图5B所示，在pH 8.5条件下孵育24 h后，EstE的相对酶活性为95%；继续孵育至100 h，EstE仍保持80%以上的相对酶活性。
2.6 酯酶EstE的最适温度和热稳定性
为了测定温度对酯酶活性的影响，在pH 8.5条件下检测了不同温度(15-65 ℃)下EstE的酶活性变化。结果表明，EstE在40 ℃时表现出最大活性。在15-40 ℃范围内，EstE的相对酶活性随着温度升高而逐步增加；在40-65 ℃范围内，EstE的酶活性则随着温度升高而逐渐下降(图6A)。为了评估EstE的热稳定性，分别将EstE置于40 ℃和100 ℃的金属浴中进行热处理，并每隔一段时间测量酶活性。结果显示，在40 ℃温育22 h后，EstE的相对酶活性为90%；在100 ℃水浴中处理4 h后，EstE仍保留50%以上的相对酶活性。热失活动力学计算表明，EstE在40 ℃和100 ℃条件下的半衰期分别为156.11 h和2.67 h，表明EstE具有优异的热稳定性(图7A、7B)。
2.7 不同添加剂对EstE酶活性的影响
在pH 8.5、25 ℃条件下，研究了不同浓度的金属离子、有机溶剂及去垢剂对酯酶EstE酶活性的影响。结果表明：(1) 金属离子中，K+、Ca2+和Ba2+对EstE具有激活作用，分别为119.44% (1 mmol/L)和167.69% (10 mmol/L)、140.94% (1 mmol/L)和155.99% (10 mmol/L)、121.20% (1 mmol/L)和153.80% (10 mmol/L)；Mg2+和Mn2+对酶活性几乎无影响，而Hg2+对酶活性有强抑制作用；Zn2+、Al3+、Ni2+和Cu2+在低浓度(1 mmol/L)时激活酶活性，而在高浓度(10 mmol/L)时抑制酶活性(图8A)；(2) 低浓度去垢剂(0.1%)对EstE酶活性具有激活作用，而高浓度去垢剂对EstE酶活性具有抑制作用，其中EstE对Tween-20的耐受性显著，相对酶活性分别为115.56% (0.1%)和85.48% (1%) (图8B)；(3) EstE对有机溶剂DMF、DMSO和甲醇的耐受性显著，在DMF (15%和30%)的存在时，相对酶活性分别为107.86%和68.99%；在甲醇(15%和30%)中，相对酶活性分别为91.58%和68.44%；DMSO对酶活性几乎无影响，相对酶活性分别为103.65%和98.67% (图8C)。
3 讨论与结论
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本研究通过对来源于灰色链霉菌的酯酶基因estE进行人工优化成功实现了其在大肠杆菌中的异源表达，并采用生物信息学分析和对硝基苯酚法对其家族分类、结构模拟及酶学性质进行了系统鉴定。在酯酶异源表达方面，通过降低G+C含量和以大肠杆菌偏好性密码子替代链霉菌密码子的优化策略，对编码酯酶基因碱基的序列进行优化，成功解决了链霉菌次级代谢产物酯酶在大肠杆菌中表达困难的问题，为链霉菌酯酶的功能研究提供了有力的工具和平台。系统发育树分析表明，酯酶EstE属于GDS(L) Family (Family II)家族。在三维结构方面，尽管酯酶EstE的核心具有α/β水解酶折叠特性，但与典型的由8条平行β-链组成的β-片核心折叠不同，EstE的中心β-片仅由7条β-链组成，这一特征与来自枯草芽孢杆菌的LipA和变铅青链霉菌TK24的EstA相似[14-15]。进一步将酯酶EstE与其他同族酯酶的生化特性进行比较(表2)，在底物特异性方面，Est804、Lip29和Est29偏好水解长链底物，而EstE与AaSGNH1和Ali5相似，偏向于水解短链底物；在比酶活方面，EstE的比酶活最高，为61.03 U/mg，高于该家族其他成员；在最适温度方面，EstE在40 ℃时表现出最大酶活性，与Ali5和Est804相似；在最适pH方面，大多数GDS(L)家族的酯酶最适pH偏碱性，EstE在pH 8.5时活性最佳[19]。
在嗜热酶挖掘方面，目前研究人员主要集中在从嗜热细菌中寻找和鉴定新型耐热酯酶作为潜在的商业生物催化剂[20-21]。近年来，研究发现一些中温菌来源的酯酶也具有出色的热稳定性。例如，Yuan等分离鉴定的来源于海洋链霉菌W007的酯酶MAS1在60 ℃下孵育1 h后仍保持80%以上的相对活性[22]。进一步地，Wang等[2]、Fang等[11]、Chang等[14]发现了来自中温变铅青链霉菌TK24的3种热稳定性酯酶，在100 ℃下，EstC和EstA分别孵育8 h和6 h时均可保留50%的酶活，EstF在100 ℃的半衰期为2.86 h。本研究发现从灰色链霉菌中分离鉴定的酯酶EstE在40 ℃和100 ℃条件下的半衰期分别为156.11 h和2.67 h，进一步证明了链霉菌可以作为新型热稳定性酶的来源。在稳定性方面，酯酶EstE在pH 8.5条件下孵育24 h后，EstE相对酶活性为95%，继续孵育至100 h，仍保持80%以上的相对酶活性。EstE优异的碱稳定性和热稳定性使其在洗涤剂和皮革工业中具有潜在应用价值[14,23]。此外，EstE对多种单价和二价金属离子、不同类型的去垢剂和有机溶剂具有一定耐受性，这为EstE在精细化工、药物合成、生物修复等领域的应用提供了可能性[24]。综上所述，通过优化密码子成功克服了酯酶等次级代谢产物在大肠杆菌中表达的难题，并拓宽了嗜热酶的来源。此外，酯酶EstE展现出优异的催化效率、热稳定性、碱稳定性和有机溶剂耐受性，使其在生物修复、造纸制浆和生物燃料等复杂工业领域具有广泛的应用前景。
基金
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  • 国家自然科学基金(22376002)
  • 安徽省自然科学基金(2108085MC78)
  • 安徽省高校自然科学研究项目(KJ2020ZD07)
  • 大学生创新创业平台训练项目(202310370544)
文献
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参考文献 引证文献
排序方式：
[1]
BORNSCHEUER UT. Microbial carboxyl esterases: classification, properties and application in biocatalysis[J]. FEMS Microbiology Reviews, 2002, 26(1): 73-81.
[2]
WANG BJ, WU S, CHANG X, CHEN J, MA JX, WANG P, ZHU GP. Characterization of a novel hyper-thermostable and chlorpyrifos-hydrolyzing carboxylesterase EstC: a representative of the new esterase family XIX[J]. Pesticide Biochemistry and Physiology, 2020, 170: 104704.
[3]
JOHAN UUM, RAHMAN RNZRA, KAMARUDIN NHA, ALI MSM. An integrated overview of bacterial carboxylesterase: structure, function and biocatalytic applications[J]. Colloids and Surfaces B: Biointerfaces, 2021, 205: 111882.
[4]
CHANDRA P, ENESPA, SINGH R, ARORA PK. Microbial lipases and their industrial applications: a comprehensive review[J]. Microbial Cell Factories, 2020, 19(1): 169.
[5]
ARPIGNY JL, JAEGER KE. Bacterial lipolytic enzymes: classification and properties[J]. Biochemical Journal, 1999, 343(Pt 1): 177-183.
[6]
LIU XY, ZHOU MY, SUN R, XING S, WU T, HE HL, CHEN JB, BIELICKI JK. Characterization of a novel esterase Est33 from an antarctic bacterium: a representative of a new esterase family[J]. Frontiers in Microbiology, 2022, 13: 855658.
[7]
KOVACIC F, BABIC N, KRAUSS U, JAEGER KE. Classification of lipolytic enzymes from bacteria[M]//ROJO F eds. Aerobic Utilization of Hydrocarbons, Oils and Lipids. Handbook of Hydrocarbon and Lipid Microbiology. Cham: Springer International Publishing, 2019: 1-35.
[8]
WU H, CHEN QM, ZHANG WL, MU WM. Overview of strategies for developing high thermostability industrial enzymes: discovery, mechanism, modification and challenges[J]. Critical Reviews in Food Science and Nutrition, 2023, 63(14): 2057-2073.
[9]
SUJITHA P, SHANTHI C. Importance of enzyme specificity and stability for the application of proteases in greener industrial processing-a review[J]. Journal of Cleaner Production, 2023, 425: 138915.
[10]
MARCHETTI A, ORLANDO M, MANGIAGALLI M, LOTTI M. A cold-active esterase enhances mesophilic properties through Mn2+ binding[J]. The FEBS Journal, 2023, 290(9): 2394-2411.
[11]
FANG JX, AN LH, YU J, MA JX, ZHOU RJ, WANG BJ. Characterization of a novel carboxylesterase from Streptomyces lividans TK24 and site-directed mutagenesis for its thermostability[J]. Journal of Bioscience and Bioengineering, 2024, 138(3): 181-187.
[12]
KUMAR A, SHARMA M, BHARDWAJ PK, VATS SK, SINGH D, KUMAR S. Copper, zinc superoxide dismutase from Caragana jubata: a thermostable enzyme that functions under a broad pH and temperature window[J]. Process Biochemistry, 2016, 51(10): 1434-1444.
[13]
AKOH CC, LEE GC, LIAW YC, HUANG TH, SHAW JF. GDSL family of serine esterases/lipases[J]. Progress in Lipid Research, 2004, 43(6): 534-552.
[14]
CHANG X, WU S, CHEN J, XIONG SQ, WANG P, SHI XQ, WANG A, WANG BJ. Characterization of a carboxylesterase with hyper-thermostability and alkali-stability from Streptomyces lividans TK24[J]. Extremophiles, 2021, 25(2): 115-128.
[15]
VAN POUDEROYEN G, EGGERT T, JAEGER KE, DIJKSTRA BW. The crystal structure of Bacillus subtilis lipase: a minimal α/β hydrolase fold enzyme[J]. Journal of Molecular Biology, 2001, 309(1): 215-226.
[16]
KNEPP ZJ, GHANER A, ROOT KT. Purification and refolding protocol for cold-active recombinant esterase Aa SGNH1 from Aphanizomenon flos-aquae expressed as insoluble inclusion bodies[J]. Preparative Biochemistry & Biotechnology, 2022, 52(4): 394-403.
[17]
HONG LG, JIAN SL, HUO YY, CHENG H, HU XJ, LI JX, CUI HL, XU XW. A novel SGNH family hydrolase Ali5 with thioesterase activity and a GNSL motif but without a classic GDSL motif from Altererythrobacter ishigakiensis [J]. Biotechnology Letters, 2019, 41(4/5): 591-604.
[18]
CHEN CH, YU G, GUO ZY, YANG QH, SU WF, XIE QF, YANG GD, REN YF, LI H. Expression, characterization, fermentation, immobilization, and application of a novel esterase Est804 from metagenomic library in pesticide degradation[J]. Frontiers in Microbiology, 2022, 13: 922506.
[19]
JO E, KIM J, LEE A, MOON K, CHA J. Identification and characterization of a novel thermostable GDSL-type lipase from Geobacillus thermocatenulatus [J]. Journal of Microbiology and Biotechnology, 2021, 31(3): 483-491.
[20]
GULERIA S, JAIN R, SINGH D, KUMAR S. A thermostable Fe/Mn SOD of Geobacillus sp. PCH100 isolated from glacial soil of Indian trans-Himalaya exhibits activity in the presence of common inhibitors[J]. International Journal of Biological Macromolecules, 2021, 179: 576-585.
[21]
AKRAM F, HAQ IU. Overexpression and characterization of TnCel12B, a hyperthermophilic GH12 endo-1,4-β-glucanase cloned from Thermotoga naphthophila RKU-10T [J]. Analytical Biochemistry, 2020, 599: 113741.
[22]
YUAN DJ, LAN DM, XIN RP, YANG B, WANG YH. Screening and characterization of a thermostable lipase from marine Streptomyces sp. strain W007[J]. Biotechnology and Applied Biochemistry, 2016, 63(1): 41-50.
[23]
RODRÍGUEZ-MEJÍA JL, HIDALGO-MANZANO IA, MURIEL-MILLÁN LF, RIVERA-GOMEZ N, SAHONERO-CANAVESI DX, CASTILLO E, PARDO-LÓPEZ L. A novel thermo-alkaline stable GDSL/SGNH esterase with broad substrate specificity from a deep-sea Pseudomonas sp.[J]. Marine Biotechnology, 2024, 26(3): 447-459.
[24]
RAFEEQ H, HUSSAIN A, SHABBIR S, ALI S, BILAL M, SHER F, IQBAL HMN. Esterases as emerging biocatalysts: mechanistic insights, genomic and metagenomic, immobilization, and biotechnological applications[J]. Biotechnology and Applied Biochemistry, 2022, 69(5): 2176-2194.
2025年第65卷第5期
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doi: 10.13343/j.cnki.wsxb.20240718
  • 接收时间：2024-11-13
  • 首发时间：2026-02-05
  • 出版时间：2025-05-04
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  • 收稿日期：2024-11-13
  • 录用日期：2025-02-21
基金
National Natural Science Foundation of China(22376002)
国家自然科学基金(22376002)
Anhui Provincial Natural Science Foundation(2108085MC78)
安徽省自然科学基金(2108085MC78)
Natural Science Research Project of Anhui Provincial Universities(KJ2020ZD07)
安徽省高校自然科学研究项目(KJ2020ZD07)
Innovation and Entrepreneurship Training Program for College Students(202310370544)
大学生创新创业平台训练项目(202310370544)
作者信息
    1.安徽师范大学 生命科学学院，安徽 芜湖
    2.分子酶学与重大代谢性疾病机理研究安徽省重点实验室，安徽 芜湖
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https://castjournals.cast.org.cn/joweb/wswxb/CN/10.13343/j.cnki.wsxb.20240718
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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