生物技术通报 ›› 2015, Vol. 31 ›› Issue (10): 171-176.doi: 10.13560/j.cnki.biotech.bull.1985.2015.10.027

• 研究报告 • 上一篇    下一篇

产脂肪酶嗜酸性真菌的筛选、鉴定与产酶条件优化

赵军,赵淑琴,杨孝朴,刘晓丽   

  1. 甘肃农业大学动物医学院, 兰州 730070
  • 收稿日期:2015-01-27 出版日期:2015-10-28 发布日期:2015-10-28
  • 作者简介:赵军, 男, 硕士, 研究方向:分子生物学与生物化学; E-mail:956015614@qq.com
  • 基金资助:
    甘肃省财政厅项目(2013NY-18)

Screening and Identification of Eosinophilic Fungal Strain Producing Lipase and Optimization of Its Lipase-producing Condition

Zhao Jun, Zhao Shuqin, Yang Xiaopu, Liu Xiaoli   

  1. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070
  • Received:2015-01-27 Published:2015-10-28 Online:2015-10-28

摘要: 旨在筛选获得高产脂肪酶菌株, 并鉴定其种属及确定最佳产酶条件。从兰州城区附近采集富含油脂的土壤, 通过维多利亚蓝与罗丹明B培养基进行菌株分离初筛。用改进铜皂-分光光度法测定酶活力值进行复筛, 获得一株产酶能力较强的菌株LZ-6。经形态学观察和分子生物学鉴定, 该菌株为雅致放射毛霉(Actinomucor elegans)。经单因素实验与正交实验确定该菌株的最佳产酶条件为:酵母粉1%、橄榄油2%、MgSO4·7H2O 0.05%、(NH42SO4 0.1%、K2HPO4 0.2%、温度30℃、pH5.0、接种量为2%、发酵5 d, 在此条件下, 产酶活力达13.35 U/mL, 比优化前提高了1.78倍。

关键词: 真菌, 脂肪酶, 筛选, 鉴定, 优化

Abstract: This experiment was devoted to screen the strains of high-yield lipase, identify the genera, and define the optimal fermentation conditions. The strains were preliminary screened from greasy soil nearby Lanzhou urban area through Victoria blue and rhodamine B plates. The strains were re-screened through a copper soap spectrophotography measuring lipase activity while using emulsified olive oil as the substrate, finally a fungal strain LZ-6 with high-yield lipase was obtained. It was identified as Actinomucor elegans through morphological observation and molecular identification. The fermentation conditions for lipase production optimized by the single factor experiments and the orthogonal experiments with strain LZ-6 were:yeast powder 1%, olive oil 2%, 0.05% MgSO4·7H2O, 0.1%(NH42SO4, 0.2% K2HPO4, fermentation temperature at 30℃, pH5.0, 2% inoculums volume, and fermentation time of 5 days; under the optimal condition, the lipase activity of strain LZ-6 reached 13.35 U/mL, it increased 1.78 times as before optimization.

Key words: fungi strain, lipase, screening, identification, optimization