枯草芽孢杆菌DC-1产维生素K_(2)的发酵条件优化  

Optimization of Fermentation Conditions for Vitamin K_(2)Production by Bacillus subtilis DC⁃1

作　　者：黄友美 曹盛 孙一斐 李诗敏 夏雪雪 袁干军[1,2] HUANG Youmei;CAO Sheng;SUN Yifei;LI Shimin;XIA Xuexue;YUAN Ganjun(Biotechnological Engineering Center for Pharmaceutical Research and Development,Jiangxi Agricultural University,Nanchang 330045,Jiangxi,China;College of Bioscience and Bioengineering,Jiangxi Agricultural University,Nanchang 330045,Jiangxi,China)

机构地区：[1]江西农业大学生物技术药物研发工程中心,江西南昌330045 [2]江西农业大学生物科学与工程学院,江西南昌330045

出　　处：《食品研究与开发》2024年第3期123-130,共8页Food Research and Development

基　　金：国家自然科学基金项目(81960636)。

摘　　要：为提高微生物发酵法生产维生素K_(2)的产量和多样性,对枯草芽孢杆菌DC-1的发酵条件进行优化。采用单因素试验对其发酵条件包括碳源、氮源和无机盐进行初步优化,通过Plackett-Burman(PB)试验筛选出对DC-1产维生素K_(2)具有显著性影响的因素,应用响应面Box-Behnken设计优化获得DC-1的最优发酵条件。结果表明,DC-1产维生素K_(2)的最优发酵条件为甘油添加量36.5 g/L、糖蜜添加量31.8 g/L、黄豆粉添加量32.5 g/L、KH2PO_(4)浓度0.3 g/100 mL、K_(2)HPO_(4)·3H_(2)O浓度0.8 g/100 mL、Mg SO_(4)·7H_(2)O浓度0.7 g/100 mL、Na Cl浓度0.5 g/100 mL。在该条件下,MK-7产量达到94.66 mg/L,比初始发酵条件提高了48.04%;同时菌株DC-1产维生素K_(2)的多样性增加,经液相色谱-质谱联用(liquid chromatograph mass spectrometer,LC-MS)鉴定能同时产5种MK同系物,分别为MK-2、MK-3、MK-4和MK-7,其中一个类似物未能在高分辨率质谱中找到,根据保留时间推测其为MK-5或MK-6,可为维生素K_(2)的工业化生产和探索维生素K_(2)同系物的生物活性奠定基础。In order to improve the yield and diversity of vitamin K_(2)produced by microbial fermentation,the fermentation conditions of Bacillus subtilis DC-1 were optimized.The fermentation conditions including carbon source,nitrogen source,and inorganic salt were optimized by single factor experiment.The factors that had a significant influence on vitamin K_(2)produced by DC-1 were screened by the Plackett-Burman(PB)experi-ment,and the optimal fermentation conditions of DC-1 were obtained by response surface Box-Behnken de-sign.The results showed that the optimal fermentation conditions for vitamin K_(2)produced by DC-1 were glyc-erol of 36.5 g/L,molasses of 31.8 g/L,soybean flour of 32.5 g/L,KH2PO_(4)of 0.3 g/100 mL,K_(2)HPO_(4)·3H_(2)O of 0.8 g/100 mL,MgSO_(4)·7H_(2)O of 0.7 g/100 mL,and NaCl of 0.5 g/100 mL.Under these conditions,the yield of MK-7 reached 94.66 mg/L,which was 48.04%higher than the initial fermentation conditions.Meanwhile,the diversity of vitamin K_(2)produced by strain DC-1 increased,and a liquid chromatograph mass spectrometer(LC-MS)identified that it could produce five MK homologues simultaneously,namely MK-2,MK-3,MK-4,and MK-7,of which one analogue could not be found in the high-resolution mass spectrometry and was presumed to be MK-5 or MK-6 based on the retention time.This could lay a foundation for the industrial production of vi-tamin K_(2)and the exploration of the biological activity of vitamin K_(2)homologues.

关 键 词：维生素K_(2) 枯草芽孢杆菌 高效液相色谱法 发酵优化 产物多样性 

分 类 号：TS201.24[轻工技术与工程—食品科学] TS201.3[轻工技术与工程—食品科学与工程]