食品科学 ›› 2018, Vol. 39 ›› Issue (14): 185-191.doi: 10.7506/spkx1002-6630-201814028

• 生物工程 • 上一篇    下一篇

高产纳豆激酶菌株的分离与发酵纳豆过程中生物胺的变化

高泽鑫1,何腊平1,2,*,刘亚兵1,高冰3,李翠芹4,刘涵玉1   

  1. (1.贵州大学酿酒与食品工程学院,贵州?贵阳 550025;2.贵州省农畜产品贮藏与加工重点实验室,贵州?贵阳 550025;3.湖北工业大学生物工程与食品学院,湖北?武汉 430068;4.贵州大学化学与化工学院,贵州?贵阳 550025)
  • 出版日期:2018-07-25 发布日期:2018-07-16
  • 基金资助:
    贵州省重大专项(黔科合重大专项字[2015]6004-5号);贵州省农业攻关项目(黔科合支撑[2016]2580号)

Screening of a High-Yield Nattokinase-Producing Strain and Changes of Biogenic Amines during Natto Fermentation

GAO Zexin1, HE Laping1,2,*, LIU Yabing1, GAO Bing3, LI Cuiqin4, LIU Hanyu1   

  1. (1. School of Liquor and Food Engineering, Guizhou University, Guiyang 550025, China;2. Key Laboratory of Agricultural and Animal Products Store & Processing of Guizhou Province, Guiyang 550025, China; 3. School of Food and Biological Engineering, Hubei University of Technology, Wuhan 430068, China; 4. School of Chemistry and Chemical Engineering, Guizhou University, Guiyang 550025, China)
  • Online:2018-07-25 Published:2018-07-16

摘要: 采用纤溶活性筛选法从贵州织金农家土制烟熏腊肉中分离到一株高产纳豆激酶菌株GULR06,对菌株的菌株形态、生理生化特征、16S rDNA序列分析以及系统发育分析,鉴定该菌株GULR06为枯草芽孢杆菌。以黄豆为主要原料进行固态发酵制备纳豆,通过高效液相色谱法对发酵过程中的生物胺进行检测,同时考察了发酵过程中纳豆激酶活力和生物量的变化。实验表明:尸胺、腐胺、2-苯乙胺、色胺、精胺、酪胺、亚精胺、组胺都能得到很好的分离,且线性关系良好,R2均大于0.994?0,相对标准偏差均小于4%。在纳豆发酵过程中生物胺总量范围在26.47~72.97?mg/kg之间,对人体不构成任何威胁。54?h时,纳豆激酶的酶活力达到最高,为(5?439.5±149)IU/g、活菌数为13(lg(CFU/g)),此时生物胺总量为(56.18±4.94)mg/kg。

关键词: 菌株GULR06, 分离, 纳豆激酶, 固态发酵, 生物量, 生物胺

Abstract: A high-yield nattokinase-producing strain, designated as GULR06, was isolated from smoked cured meat produced by local farmers in Zhijin, Guizhou by measuring fibrinolytic activity of the enzyme produced by this strain. The strain GULR06 was identified as Bacillus subtilis based on morphology, physiological and biochemical characteristics, 16S rDNA sequence analysis and phylogenetic analysis. Natto was prepared by solid state fermentation with soybean as the major ingredient, and the changes of biogenic amines during the fermentation process were detected by high performance liquid chromatography (HPLC). The changes of nattokinase activity and biomass were also investigated. The results showed that cadaverine, putrescine, 2-phenethylamine, tryptamine, spermine, tyramine, spermidine and histamine were well separated by HPLC. The method showed good linearity with R2 greater than 0.994 0, and relative standard derivation (RSD) lower than 4%. The total content of biogenic amine during natto fermentation was in the range of 26.47–72.97 mg/kg, which was safe for human body. After fermentation for 54 h, the activity of nattokinase, (5 439.5 ± 149) IU/g, was the highest, the number of viable cells was 13 (lg(CFU/g)), and the total amount of biogenic amines was (56.18 ± 4.94) mg/kg.

Key words: GULR06 strain, isolate, nattokinase, solid state fermentation, biomass, biogenic amines

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