云南高原湖泊抚仙湖和星云湖的酵母菌胞外酶活性
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摘要
【背景】高原湖泊因其海拔高、气压低、辐射强、氧气含量低,是一类特殊环境,而其中的微生物是高原湖泊生态系统物质循环与能量流动的重要参与者,其胞外酶活性的表现决定其适应这一特殊环境的方式与能力。【目的】对分离自云南高原湖泊抚仙湖和星云湖湖水的酵母菌进行产胞外酶活性的筛选,以期获得具有潜在应用价值的活性菌株。【方法】在5°C和25°C培养温度下,采用平板筛选法对两个湖泊酵母菌进行产胞外蛋白酶、纤维素酶、淀粉酶、脂肪酶、几丁质酶、木聚糖酶、植酸酶、菊粉酶、漆酶、锰依赖过氧化物酶和木质素过氧化物酶活性的筛选。【结果】抚仙湖和星云湖的所有测试酵母菌菌株至少都能产1种胞外酶,且主要产植酸酶、菊粉酶和淀粉酶;其次为脂肪酶、纤维素酶、木聚糖酶、锰依赖过氧化物酶和木质素过氧化物酶;产几丁质酶、蛋白酶和漆酶的酵母菌很少,星云湖酵母菌都不产漆酶。培养温度为5°C时,抚仙湖和星云湖的酵母菌产5种及5种以上胞外酶的活性菌株数均多于25°C。【结论】抚仙湖和星云湖的酵母菌产胞外酶菌株多样性丰富,胞外酶种类多样,产酶酵母菌可能参与高原湖泊生态系统的物质循环;筛选得到的产胞外酶菌株为开发与利用高原湖泊酶资源提供了良好的种质资源,具有进一步研究的价值。
[Background] Plateau lakes are a type of special environments due to the conditions of high altitude, low air pressure, high ultraviolet radiation and lower oxygen content; while microorganisms in the plateau lakes are important participants in substance circulation and energy flow of lake ecosystems, and the situations/performances of their extracellular enzymatic activities formulate their adaptation ways and abilities to the special environment. [Objective] To screen yeasts with extracellular enzymatic activities, which were isolated from Fuxian Lake and Xingyun Lake in Yunnan Plateau, and to obtain active yeasts with potential application values. [Methods]Yeasts exhibiting activities of extracellular proteinase, cellulase, amylase, lipase, chitinase, xylanase,phytase, inulase, laccase, manganese-dependent peroxidase and lignin peroxidase were detected by using screening plate method at 5 °C and 25 °C. [Results] All yeast strains exhibited at least one extracellular enzymatic activity, mainly exhibited activities of phytase, inulase and amylase, secondly exhibited activities of lipase, cellulase, xylanas and manganese-dependent peroxidase and lignin peroxidase. The number of strains exhibited activities of chitinase, proteinase and laccase activities were lower, and the yeast strains isolated from Xingyun Lake did not exhibited laccase activity. The number of yeast strains exhibited at least five extracellular enzymatic activities was higher at 5 °C than that at 25 °C. [Conclusion] The rich diversity of yeasts with extracellular enzymatic activities exists in the two lakes, and hence these yeasts might participate in the circulation of substances in the plateau lake ecosystems. For further exploitation and utilization of extracellular enzymes, these active yeasts can be used as the resources of extracellular enzymes, and it is worth to further explore their application potential.
[Background] Plateau lakes are a type of special environments due to the conditions of high altitude, low air pressure, high ultraviolet radiation and lower oxygen content; while microorganisms in the plateau lakes are important participants in substance circulation and energy flow of lake ecosystems, and the situations/performances of their extracellular enzymatic activities formulate their adaptation ways and abilities to the special environment. [Objective] To screen yeasts with extracellular enzymatic activities, which were isolated from Fuxian Lake and Xingyun Lake in Yunnan Plateau, and to obtain active yeasts with potential application values. [Methods]Yeasts exhibiting activities of extracellular proteinase, cellulase, amylase, lipase, chitinase, xylanase,phytase, inulase, laccase, manganese-dependent peroxidase and lignin peroxidase were detected by using screening plate method at 5 °C and 25 °C. [Results] All yeast strains exhibited at least one extracellular enzymatic activity, mainly exhibited activities of phytase, inulase and amylase, secondly exhibited activities of lipase, cellulase, xylanas and manganese-dependent peroxidase and lignin peroxidase. The number of strains exhibited activities of chitinase, proteinase and laccase activities were lower, and the yeast strains isolated from Xingyun Lake did not exhibited laccase activity. The number of yeast strains exhibited at least five extracellular enzymatic activities was higher at 5 °C than that at 25 °C. [Conclusion] The rich diversity of yeasts with extracellular enzymatic activities exists in the two lakes, and hence these yeasts might participate in the circulation of substances in the plateau lake ecosystems. For further exploitation and utilization of extracellular enzymes, these active yeasts can be used as the resources of extracellular enzymes, and it is worth to further explore their application potential.
引文
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[8]Wang XH,Chi ZM.Isolation and identification of amylase-producing marine yeasts[J].Periodical of Ocean University of China,2007,37(S2):95-100(in Chinese)王晓红,池振明.产淀粉酶海洋酵母菌的筛选及鉴定[J].中国海洋大学学报:自然科学版,2007,37(S2):95-100
[9]Zhang L,Chi ZM.Screening and identification of a cellulase producing marine yeast and medium fermentation condition optimization for cellulase production[J].Periodical of Ocean University of China,2007,37(S2):101-108(in Chinese)张亮,池振明.1株产纤维素酶海洋酵母菌的筛选、鉴定及发酵条件优化[J].中国海洋大学学报:自然科学版,2007,37(S2):101-108
[10]Song W,Jiang LJ,Shen AR,et al.Isolation of lipase yeast strains and their induction with ultraviolet radiation[J].Journal of Central South University of Forestry&Technology,2009,29(3):55-59(in Chinese)宋炜,蒋丽娟,申爱荣,等.高产脂肪酶酵母菌株的分离筛选及紫外诱变[J].中南林业科技大学学报,2009,29(3):55-59
[11]Lata S,Rastogi S,Kapoor A,et al.Influence of culture conditions on production of phytase by Zygosaccharomyces baili var.balil.[J].Journal of Environmental Biology,2015,36(4):947-954
[12]Hui FL,Feng JR,Yang KJ,et al.Screening and identification of chitinase-producing yeast[J].Journal of Northeast Forestry University,2007,35(8):66-67(in Chinese)惠丰立,冯金荣,杨柯金,等.产几丁质酶酵母菌的筛选与鉴定[J].东北林业大学学报,2007,35(8):66-67
[13]Wang GY,Song AR,Chen GJ.Screening and identification of yeast strains producing inulinases[J].Journal of Microbiology,2006,26(4):39-41(in Chinese)王光远,宋爱荣,陈冠军.产菊粉酶酵母菌株的筛选及菌种鉴定[J].微生物学杂志,2006,26(4):39-41
[14]Zheng XB,Guo LQ,Fu XY,et al.Optimization of laccase production conditions of a newly isolated Trichoderma spp.strain La Tr01 from southern China[J].Chinese Journal of Bioprocess Engineering,2007,5(4):19-24(in Chinese)郑晓冰,郭丽琼,付小燕,等.木霉La Tr 01菌株产漆酶发酵的条件[J].生物加工过程,2007,5(4):19-24
[15]Cai L,Yin JF,Yang LP,et al.Several qualitative methods for the screening of fungi to decompose lignin[J].Microbiology China,2002,29(1):67-69(in Chinese)蔡磊,尹峻峰,杨丽萍,等.几种简便的木质素降解真菌定性筛选方法[J].微生物学通报,2002,29(1):67-69
[16]Zhang JZ,Pang ZW,Liang JJ,et al.The biosynthesis of xylanase by Aureobasidium pulluans XYE-7[J].Modern Food Science&Technology,2007,23(1):12-14(in Chinese)张建珍,庞宗文,梁静娟,等.影响出芽短梗霉XYE-7木聚糖酶生物合成的条件[J].现代食品科技,2007,23(1):12-14
[17]Zhou XL,Li ZY,Yang LY,et al.Identification of yeasts isolation of Chenghai Lake,a plateau lake in Yunnan province[J].Acta Microbiologica Sinica,2011,51(4):547-553(in Chinese)周新丽,李治滢,杨丽源,等.云南程海湖酵母菌多样性及应用[J].微生物学报,2011,51(4):547-553
[18]Wang QM,Yurkov AM,G?ker M,et al.Phylogenetic classification of yeasts and related taxa within Pucciniomycotina[J].Studies in Mycology,2015,81:149-189
[19]Liu XZ,Wang QM,G?ker M,et al.Towards an integrated phylogenetic classification of the Tremellomycetes[J].Studies in Mycology,2015,81:85-147
[20]Brand?o LR,Libkind D,Vaz ABM,et al.Yeasts from an oligotrophic lake in Patagonia(Argentina):diversity,distribution and synthesis of photoprotective compounds and extracellular enzymes[J].Federation of European Microbiological Societies Microbiology Ecology,2011,76(1):1-13
[21]Yan YP,Li ZY,Dong MH,et al.Yeasts from Yangzonghai Lake in Yunnan(China):diversity and extracellular enzymes[J].Acta Microbiologica Sinica,2013,53(11):1205-1212(in Chinese)严亚萍,李治滢,董明华,等.云南阳宗海酵母菌种群结构及产胞外酶测试[J].微生物学报,2013,53(11):1205-1212
[22]Li ZY,Zhou XL,Zhou B,et al.Diversity and extracellular enzymes of yeasts from Chenghai Lake in winter[J].Mycosystema,2017,36(2):177-185(in Chinese)李治滢,周新丽,周斌,等.云南程海湖冬季酵母菌多样性及胞外酶活性研究[J].菌物学报,2017,36(2):177-185
[23]Huang RZ.Study on temporal and spatial variation,influence factors of the extracellular enzymes activity from water column and sediment in Erhai Lake[D].Nanchang:Master’s Thesis of Nanchang University,2012(in Chinese)黄睿智.洱海水体及沉积物酶活性的时空变化及其影响因素的研究[D].南昌:南昌大学硕士学位论文,2012
[24]MolnárováJ,VadkertiováR,StratilováE.Extracellular enzymatic activities and physiological profiles of yeasts colonizing fruit trees[J].Journal of Basic Microbiology,2014,54(S1):S74-S84
[25]Alcaíno J,Cifuentes V,Baeza M.Physiological adaptations of yeasts living in cold environments and their potential applications[J].World Journal of Microbiology and Biotechnology,2015,31(10):1467-1473
[26]Loperena L,Soria V,Varela H,et al.Extracellular enzymes produced by microorganisms isolated from maritime Antarctica[J].World Journal of Microbiology and Biotechnology,2012,28(5):2249-2256
[27]Chi ZM,Chi Z,Zhang T,et al.Production,characterization and gene cloning of the extracellular enzymes from the marine-derived yeasts and their potential applications[J].Biotechnology Advances,2009,27(3):236-255
[28]Brizzio S,Turchetti B,de García V,et al.Extracellular enzymaticactivities of basidiomycetous yeasts isolated from glacial and subglacial waters of northwest Patagonia(Argentina)[J].Canadian Journal of Microbiology,2007,53(4):519-525
[29]Carrasco M,Rozas JM,Barahona S,et al.Diversity and extracellular enzymatic activities of yeasts isolated from KingGeorge Island,the sub-Antarctic region[J].BMC Microbiology,2012,12:251
[30]Yang QX,Hao Z,Li XL,et al.Extracellular enzyme production and phylogenetic distribution of yeasts in wastewater treatment systems[J].Bioresource Technology,2013,129:264-273
[2]Zeng YX,Chen B.Progress and application prospects in the study on antarctic cold-adapted microorganisms[J].Chinese Journal of Polar Research,1999,11(2):143-152(in Chinese)曾胤新,陈波.南极低温微生物研究及其应用前景[J].极地研究,1999,11(2):143-152
[3]Zaky AS,Tucker GA,Daw ZY,et al.Marine yeast isolation and industrial application[J].Federation of European Microbiological Societies Yeast Research,2014,14(6):813-825
[4]Yang JL,Li J,Li JW,et al.Summarize on the research of the hydro-bios and aquatic environment in Fuxian Lake and Xingyun Lake[J].Yunnan Geographic Environment Research,2012,24(2):98-102(in Chinese)杨加林,李杰,李经纬,等.抚仙湖-星云湖水生生物与水环境研究综述[J].云南地理环境研究,2012,24(2):98-102
[5]Xu JY,Zhao XF,LüCY,et al.Characteristics of water environment and comprehensive improvement research of pollution prevention in Plateau Lake basin[J].EnvironmentalScience and Management,2015,40(3):49-52(in Chinese)许杰玉,赵晓飞,吕春英,等.高原湖泊流域水环境特征与污染防治综合整治研究[J].环境科学与管理,2015,40(3):49-52
[6]Guo XF,Li ZY,Dong MH,et al.Spatial dynamics of yeast community and its relationship to environmental factors in Lake Fuxian,Yunnan Province[J].Journal of Lake Science,2016,28(2):358-369(in Chinese)郭小芳,李治滢,董明华,等.云南高原湖泊抚仙湖酵母菌空间分布及其与环境因子的关系[J].湖泊科学,2016,28(2):358-369
[7]Chen R,Guan SS,LüGZ,et al.Preliminary screening of protease-producing Mucor strain[J].Journal of Microbiology,2008,28(1):101-104(in Chinese)陈嵘,关珊珊,吕国忠,等.产蛋白酶毛霉菌株的初步筛选[J].微生物学杂志,2008,28(1):101-104
[8]Wang XH,Chi ZM.Isolation and identification of amylase-producing marine yeasts[J].Periodical of Ocean University of China,2007,37(S2):95-100(in Chinese)王晓红,池振明.产淀粉酶海洋酵母菌的筛选及鉴定[J].中国海洋大学学报:自然科学版,2007,37(S2):95-100
[9]Zhang L,Chi ZM.Screening and identification of a cellulase producing marine yeast and medium fermentation condition optimization for cellulase production[J].Periodical of Ocean University of China,2007,37(S2):101-108(in Chinese)张亮,池振明.1株产纤维素酶海洋酵母菌的筛选、鉴定及发酵条件优化[J].中国海洋大学学报:自然科学版,2007,37(S2):101-108
[10]Song W,Jiang LJ,Shen AR,et al.Isolation of lipase yeast strains and their induction with ultraviolet radiation[J].Journal of Central South University of Forestry&Technology,2009,29(3):55-59(in Chinese)宋炜,蒋丽娟,申爱荣,等.高产脂肪酶酵母菌株的分离筛选及紫外诱变[J].中南林业科技大学学报,2009,29(3):55-59
[11]Lata S,Rastogi S,Kapoor A,et al.Influence of culture conditions on production of phytase by Zygosaccharomyces baili var.balil.[J].Journal of Environmental Biology,2015,36(4):947-954
[12]Hui FL,Feng JR,Yang KJ,et al.Screening and identification of chitinase-producing yeast[J].Journal of Northeast Forestry University,2007,35(8):66-67(in Chinese)惠丰立,冯金荣,杨柯金,等.产几丁质酶酵母菌的筛选与鉴定[J].东北林业大学学报,2007,35(8):66-67
[13]Wang GY,Song AR,Chen GJ.Screening and identification of yeast strains producing inulinases[J].Journal of Microbiology,2006,26(4):39-41(in Chinese)王光远,宋爱荣,陈冠军.产菊粉酶酵母菌株的筛选及菌种鉴定[J].微生物学杂志,2006,26(4):39-41
[14]Zheng XB,Guo LQ,Fu XY,et al.Optimization of laccase production conditions of a newly isolated Trichoderma spp.strain La Tr01 from southern China[J].Chinese Journal of Bioprocess Engineering,2007,5(4):19-24(in Chinese)郑晓冰,郭丽琼,付小燕,等.木霉La Tr 01菌株产漆酶发酵的条件[J].生物加工过程,2007,5(4):19-24
[15]Cai L,Yin JF,Yang LP,et al.Several qualitative methods for the screening of fungi to decompose lignin[J].Microbiology China,2002,29(1):67-69(in Chinese)蔡磊,尹峻峰,杨丽萍,等.几种简便的木质素降解真菌定性筛选方法[J].微生物学通报,2002,29(1):67-69
[16]Zhang JZ,Pang ZW,Liang JJ,et al.The biosynthesis of xylanase by Aureobasidium pulluans XYE-7[J].Modern Food Science&Technology,2007,23(1):12-14(in Chinese)张建珍,庞宗文,梁静娟,等.影响出芽短梗霉XYE-7木聚糖酶生物合成的条件[J].现代食品科技,2007,23(1):12-14
[17]Zhou XL,Li ZY,Yang LY,et al.Identification of yeasts isolation of Chenghai Lake,a plateau lake in Yunnan province[J].Acta Microbiologica Sinica,2011,51(4):547-553(in Chinese)周新丽,李治滢,杨丽源,等.云南程海湖酵母菌多样性及应用[J].微生物学报,2011,51(4):547-553
[18]Wang QM,Yurkov AM,G?ker M,et al.Phylogenetic classification of yeasts and related taxa within Pucciniomycotina[J].Studies in Mycology,2015,81:149-189
[19]Liu XZ,Wang QM,G?ker M,et al.Towards an integrated phylogenetic classification of the Tremellomycetes[J].Studies in Mycology,2015,81:85-147
[20]Brand?o LR,Libkind D,Vaz ABM,et al.Yeasts from an oligotrophic lake in Patagonia(Argentina):diversity,distribution and synthesis of photoprotective compounds and extracellular enzymes[J].Federation of European Microbiological Societies Microbiology Ecology,2011,76(1):1-13
[21]Yan YP,Li ZY,Dong MH,et al.Yeasts from Yangzonghai Lake in Yunnan(China):diversity and extracellular enzymes[J].Acta Microbiologica Sinica,2013,53(11):1205-1212(in Chinese)严亚萍,李治滢,董明华,等.云南阳宗海酵母菌种群结构及产胞外酶测试[J].微生物学报,2013,53(11):1205-1212
[22]Li ZY,Zhou XL,Zhou B,et al.Diversity and extracellular enzymes of yeasts from Chenghai Lake in winter[J].Mycosystema,2017,36(2):177-185(in Chinese)李治滢,周新丽,周斌,等.云南程海湖冬季酵母菌多样性及胞外酶活性研究[J].菌物学报,2017,36(2):177-185
[23]Huang RZ.Study on temporal and spatial variation,influence factors of the extracellular enzymes activity from water column and sediment in Erhai Lake[D].Nanchang:Master’s Thesis of Nanchang University,2012(in Chinese)黄睿智.洱海水体及沉积物酶活性的时空变化及其影响因素的研究[D].南昌:南昌大学硕士学位论文,2012
[24]MolnárováJ,VadkertiováR,StratilováE.Extracellular enzymatic activities and physiological profiles of yeasts colonizing fruit trees[J].Journal of Basic Microbiology,2014,54(S1):S74-S84
[25]Alcaíno J,Cifuentes V,Baeza M.Physiological adaptations of yeasts living in cold environments and their potential applications[J].World Journal of Microbiology and Biotechnology,2015,31(10):1467-1473
[26]Loperena L,Soria V,Varela H,et al.Extracellular enzymes produced by microorganisms isolated from maritime Antarctica[J].World Journal of Microbiology and Biotechnology,2012,28(5):2249-2256
[27]Chi ZM,Chi Z,Zhang T,et al.Production,characterization and gene cloning of the extracellular enzymes from the marine-derived yeasts and their potential applications[J].Biotechnology Advances,2009,27(3):236-255
[28]Brizzio S,Turchetti B,de García V,et al.Extracellular enzymaticactivities of basidiomycetous yeasts isolated from glacial and subglacial waters of northwest Patagonia(Argentina)[J].Canadian Journal of Microbiology,2007,53(4):519-525
[29]Carrasco M,Rozas JM,Barahona S,et al.Diversity and extracellular enzymatic activities of yeasts isolated from KingGeorge Island,the sub-Antarctic region[J].BMC Microbiology,2012,12:251
[30]Yang QX,Hao Z,Li XL,et al.Extracellular enzyme production and phylogenetic distribution of yeasts in wastewater treatment systems[J].Bioresource Technology,2013,129:264-273
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