圆柚酮生物转化菌种筛选及培养基组分优化
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华中农业大学

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TS201.3

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国家自然科学基金


Screening of strains for biotransformation to nootkatone and optimization of medium components
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    摘要:

    本研究从湖北省华中农业大学柑橘果园取得土样,以瓦伦西亚橘烯为唯一碳源,首先根据在最小培养基中测得的生长曲线进行初筛,选取生长良好的菌株;以初筛菌株转化瓦伦西亚橘烯生成圆柚酮的含量为指标进行复筛,选取圆柚酮转化量高的菌株;利用SPME-GC-MS技术,对筛选所得菌株转化瓦伦西亚橘烯生成的产物种类及含量进行测定与分析;通过生理生化试验以及16SrDNA测序鉴定对所筛菌株进行菌种鉴定,确定所筛菌株为伯克霍尔德菌属,将其命名为Burkholderiasp. PZQ14。在LB培养基的基础上,以圆柚酮转化量为指标,进行发酵培养基成分单因素试验,在此基础上,通过响应面优化试验,探讨培养基组分不同浓度组合对响应值圆柚酮转化量的影响。结果表明,最优培养基组分为:葡萄糖浓度为38g/L,蛋白胨浓度为25g/L,FeSO4浓度为0.60g/L。在此条件下进行3次验证试验,圆柚酮转化量为328.69±17.88mg/L,转化率为35.73±1.24%,相比LB培养基圆柚酮转化量提高了251mg/L,转化率提高了27.30%。

    Abstract:

    In this study, soil samples were obtained from citrus orchard from Huazhong Agricultural University. Using valencene as the only carbon source, the strains with good growth characteristics were firstly screened according to the growth curve measured in the minimum culture medium. And the strains with high conversion amount of nootkatone were screened again. The types and contents of the products produced by valencene biotransformation were measured and analyzed using SPME-GC-MS. Through physiological and biochemical tests and 16SrDNA sequencing, the screened strains were identified as Burkholderia sp., which was named Burkholderia sp. PZQ14. Single-factor test of fermentation medium components was conducted using LB medium as a starting point and nootkatone yield as an index. On this basis, the effect of different concentration of culture medium components on nootkatone yield was investigated by response surface optimization test. The results showed that the optimal medium components were: glucose concentration of 38 g/L, peptone concentration of 25 g/L, and FeSO4 concentration of 0.60 g/L. The content of nootkatone was 328.69 ± 17.88 mg/L and the conversion ratio was 35.73 ± 1.24% when three validation tests were conducted under these conditions. The nootkatone yield increased by 251 mg/L and the conversion ratio increased by 27.30% compared to LB medium.

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  • 收稿日期:2021-02-18
  • 最后修改日期:2021-03-22
  • 录用日期:2021-03-27
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