渤海海域低温GOD菌株筛选及鉴定
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摘要
海洋微生物来源的葡萄糖氧化酶通常具有低温或常温下高催化活性的特点,在食品及饲料工业中具有广泛的应用。笔者所在项目组利用平板显色法,从大连渤海海域的海泥和海鱼中分离产低温葡萄糖氧化酶的微生物,经摇瓶发酵法测定各菌株的产酶能力,最终通过菌株的形态学特征和生理生化特征进行初步鉴定。该实验共筛选得到了具有产低温葡萄糖氧化酶能力的菌株6株,其中细菌2株、酵母菌2株和霉菌2株。产葡萄糖氧化酶的酶活力最高的菌为编号5-7霉菌,其酶活力为0.904 U/mL;产葡萄糖氧化酶的酶活力最低的菌为编号2-4酵母菌,其酶活力为0.135 U/mL。通过形态学及生理生化特征鉴定,初步确定了具有产低温葡萄糖氧化酶能力的两株细菌的分类:细菌8-2可能为假单胞菌属,细菌8-3可能为分枝杆菌属。该研究发掘了潜在的低温葡萄糖氧化酶生产菌株,为今后葡萄糖氧化酶的工业化生产及应用奠定了前期研究基础。
Glucose oxidase(GOD), which is the source of marine microorganisms, usually has the characteristics of high catalytic activities at low temperature or normal temperature, and is widely used in the food and feed industry. In this paper,the microorganism producing low-temperature GOD was isolated from marine mud and fish in Bohai sea area of Dalian by plate chromogenic method, and the enzyme production capacity of each strain was determined by flask fermentation method.Finally, the microorganism was preliminarily identified by morphological, physiological and biochemical characteristics of the strain. Six strains with the ability to produce hypothermic glucose oxidase were screened, including 2 bacterial strains, 2 yeast strains and 2 mold strains. The bacteria with the highest GOD enzyme activity were mycetes No.5-7, and the GOD enzyme activity was 0.904 U/m L. The bacteria with the lowest GOD enzyme activity were yeast No. 2-4, whose GOD enzyme activity was 0.135 U/m L. Through the identification of morphological, physiological and biochemical characteristics, the classification of two strains of bacteria with god-producing ability was preliminarily determined: bacteria 8-2 may be pseudomonas and bacteria 8-3 may be mycobacterium. In this study, potential low-temperature GOD production strains were discovered, which laid a foundation for the future research of industrial production and application of GOD.
Glucose oxidase(GOD), which is the source of marine microorganisms, usually has the characteristics of high catalytic activities at low temperature or normal temperature, and is widely used in the food and feed industry. In this paper,the microorganism producing low-temperature GOD was isolated from marine mud and fish in Bohai sea area of Dalian by plate chromogenic method, and the enzyme production capacity of each strain was determined by flask fermentation method.Finally, the microorganism was preliminarily identified by morphological, physiological and biochemical characteristics of the strain. Six strains with the ability to produce hypothermic glucose oxidase were screened, including 2 bacterial strains, 2 yeast strains and 2 mold strains. The bacteria with the highest GOD enzyme activity were mycetes No.5-7, and the GOD enzyme activity was 0.904 U/m L. The bacteria with the lowest GOD enzyme activity were yeast No. 2-4, whose GOD enzyme activity was 0.135 U/m L. Through the identification of morphological, physiological and biochemical characteristics, the classification of two strains of bacteria with god-producing ability was preliminarily determined: bacteria 8-2 may be pseudomonas and bacteria 8-3 may be mycobacterium. In this study, potential low-temperature GOD production strains were discovered, which laid a foundation for the future research of industrial production and application of GOD.
引文
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[13]朱运平,褚文丹,李秀婷,等.1株产胞外GOD黑曲霉的液体发酵条件优化[J].中国食品学报,2014,14(5):90-98.
[14]孙悦民.中国海洋资源开发现状及对策[J].海洋信息,2009(3):20-23.
[15]王芳,栾维新.我国海洋资源开发活动中存在的问题与建议[J].中国人口·资源与环境,2001(S2):33-35.
[16]王海英.资源综论海洋资源开发与海洋产业结构发展重点与方向[J].海洋开发与管理,2002,19(4):23-28.
[17]石淑钰,张庆芳,迟乃玉,等.一株海洋低温GOD菌株的筛选、鉴定及部分酶学性质[J].微生物学通报,2014,41(5):832-838.
[18]叶日英,徐德峰,孙力军,等.海洋源高产GOD细菌的筛选和主要酶学性质[J].浙江农业学报,2018,30(4):672-678.
[19]肖志明.葡萄糖氧化酶的应用和检测方法研究进展[J].中国畜牧杂志,2014,50(18):76-81.(责任编辑邸开宇)
[2]段钢.工业酶近期发展情况与展望[J].生物产业技术,2017(1):7-12.
[3]FOULDS N C,LOWE C R.Immobilization of glucose oxidase in ferrocene-modified pyrrole polymers[J].Analytical Chemistry,1988,60(22):2473.
[4]KANG X,WANG J,WU H,et al.Glucose oxidase-graphene-chitosan modified electrode for direct electrochemistry and glucose sensing[J].Biosensors&Bioelectronics,2009,25(4):901-905.
[5]NAKAMURA S,FUJIKI S.Comparative studies on the glucose oxidases of Aspergillus niger and Penicillium amagasakiense[J].Biochemical Journal,1968,63(1):51-58.
[6]WITT S,SINGH M,KALISZ H,et al.Structural and kinetic properties of nonglycosylated recombinant Penicillium amagasakiense glucose oxidase expressed in Escherichia coli[J].Applied and Environmental Microbiology,1998,64(4):1405-1411.
[7]熊云霞.葡萄糖氧化酶在畜牧业中的应用及检测方法研究进展[J].饲料工业,2016,37(4):15-20.
[8]刘志国,张志东,冯定远.日粮中添加过氧化氢酶对断奶仔猪生长性能、肠道形态及抗氧化性能的影响[J].中国饲料,2017(1):23-27.
[9]孙灏,王洪恩.硫醇/金自组装膜上组装葡萄糖氧化酶及其应用[J].济宁医学院学报,1999(1):4-7.
[10]吴国权,刘仲明,李忠彦.碳纳米管和纳米金自组装固定GOD的葡萄糖传感器的研究[J].医疗卫生装备,2009,30(4):4-7.
[11]吴迪,吴健.金-硫键自组装生物分子膜[J].化学通报,2004,67(2):132-137.
[12]王政,张大伟.抗生素替代技术在畜牧业中的应用[J].中国饲料,2017(24):30-34.
[13]朱运平,褚文丹,李秀婷,等.1株产胞外GOD黑曲霉的液体发酵条件优化[J].中国食品学报,2014,14(5):90-98.
[14]孙悦民.中国海洋资源开发现状及对策[J].海洋信息,2009(3):20-23.
[15]王芳,栾维新.我国海洋资源开发活动中存在的问题与建议[J].中国人口·资源与环境,2001(S2):33-35.
[16]王海英.资源综论海洋资源开发与海洋产业结构发展重点与方向[J].海洋开发与管理,2002,19(4):23-28.
[17]石淑钰,张庆芳,迟乃玉,等.一株海洋低温GOD菌株的筛选、鉴定及部分酶学性质[J].微生物学通报,2014,41(5):832-838.
[18]叶日英,徐德峰,孙力军,等.海洋源高产GOD细菌的筛选和主要酶学性质[J].浙江农业学报,2018,30(4):672-678.
[19]肖志明.葡萄糖氧化酶的应用和检测方法研究进展[J].中国畜牧杂志,2014,50(18):76-81.(责任编辑邸开宇)
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