食品科学 ›› 2022, Vol. 43 ›› Issue (4): 88-94.doi: 10.7506/spkx1002-6630-20210113-145

• 生物工程 • 上一篇    下一篇

指状青霉DSM62840生物转化柠檬烯生产α-松油醇过程中关键酶的分离纯化及质谱鉴定

张璐璐,范刚,李晓,任婧楠,潘思轶,黄文,何进   

  1. (1.河南工业大学粮油食品学院,河南 郑州 450001;2.华中农业大学食品科学技术学院,环境食品学教育部重点实验室,湖北 武汉 430070;3.华中农业大学生命科学技术学院,湖北 武汉 430070)
  • 出版日期:2022-02-25 发布日期:2022-03-08
  • 基金资助:
    国家自然科学基金面上项目(31671824);国家自然科学基金青年科学基金项目(32101868)

Isolation, Purification and Mass Spectrometry Identification of Enzyme Involved in Limonene Biotransformation into α-Terpineol by Penicillium digitatum DSM62840

ZHANG Lulu, FAN Gang, LI Xiao, REN Jingnan, PAN Siyi, HUANG Wen, HE Jin   

  1. (1. College of Food Science and Engineering, Henan University of Technology, Zhengzhou 450001, China; 2. Key Laboratory of Environment Correlative Dietology, Ministry of Education, College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China; 3. College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China)
  • Online:2022-02-25 Published:2022-03-08

摘要: 以指状青霉DSM62840为研究对象,通过高压法破碎细胞,采用超高速离心、硫酸铵沉淀、羟基磷灰石层析和凝胶过滤层析等方法对指状青霉DSM62840生物转化柠檬烯生产α-松油醇过程中的关键酶进行分离纯化。经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳检测,结果显示在分子质量约为50 kDa处有一条明显的蛋白条带,并通过液相色谱-串联质谱分析初步确定该条带中的蛋白可能是二氢硫辛酰胺脱氢酶(PDIDSM_08010)。本研究为α-松油醇类香料的合成提供理论依据和参考。

关键词: 指状青霉DSM62840;柠檬烯微生物转化;α-松油醇;分离纯化;质谱

Abstract: The key enzyme involved in the biotransformation of limonene into α-terpineol by Penicillium digitatum DSM62840 was extracted and purified by sequential ultracentrifugation, ammonium sulfate precipitation, hydroxyapatite chromatography and gel filtration chromatography after high pressure disruption of cells. The results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed an obvious protein band with a molecular mass of approximatelty 50 kDa. This protein was tentatively identified as dihydrolipoyl dehydrogenase (PDIDSM_08010) by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results of this study may provide a basis and reference for α-terpineol production.

Key words: Penicillium digitatum DSM62840; limonene biotransformation; α-terpineol; separation and purification; mass spectrometry

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