食品科学

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北极海洋红球菌(Rhodococcus sp.)B7740产类胡萝卜素的提取条件优化及甲基萘醌类类胡萝卜素鉴定

陈亚淑1,汪 荣1,谢笔钧1,杨季芳2,陈吉刚2,孙智达1,*   

  1. 1.华中农业大学食品科学技术学院,湖北 武汉 430070;2.浙江万里学院生物与环境学院,浙江 宁波 315100
  • 出版日期:2016-01-25 发布日期:2016-01-15
  • 通讯作者: 孙智达
  • 基金资助:

    公益性行业(海洋)科研专项(201405015-5)

Optimization of Extraction of Carotenoids and Identification of Menaquinone Carotenoids from Rhodococcus sp. B7740

CHEN Yashu1, WANG Rong1, XIE Bijun1, YANG Jifang2, CHEN Jigang2, SUN Zhida1,*   

  1. 1. College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China;
    2. College of Biological and Environmental Science, Zhejiang Wanli University, Ningbo 315100, China
  • Online:2016-01-25 Published:2016-01-15
  • Contact: SUN Zhida

摘要:

利用酶的高效性和专一性破裂细菌细胞壁,实现对类胡萝卜素的温和提取并鉴定。以加酶量、酶解时间和料液比作为单因素进行正交试验,得出最优提取条件为:菌粉与溶菌酶配比1∶10(mg/mL)、酶解时间80 min、料液比1∶80(mg/mL),此条件下类胡萝卜素提取量为675 μg/g。采用液相色谱-质谱串联进行类胡萝卜素的结构鉴定,液相色谱-质谱条件为:色谱柱YMCC30(4.6 mm×150 mm,5 μm)、检测波长450 nm、大气压力化学电离源、正离子模式、离子源温度230 ℃、充电电压2 000 V、质荷比扫描范围200~1 200,鉴定出分子式分别为C51H76O2和C51H74O2的2 种甲基萘醌类胡萝卜素。

关键词: 类胡萝卜素, 酶法提取, 高效液相色谱, 液相色谱-质谱, 红球菌B7740

Abstract:

The mild extraction of carotenoids from Rhodococcus sp. using lysozyme, which can break down the bacterial
cell wall efficiently and specifically, was achieved. An orthogonal array design was employed to determine the optimal
hydrolysis conditions as follows: bacterial cell powder/lysozyme ratio, 1:10 (mg/mL); hydrolysis time, 80 min; and solid/
liquid ratio, 1:80 (mg/mL). The extracted carotenoids were identified by liquid chromatography-mass spectrometry (LC-MS).
The samples were separated on a YMCC30 column (4.6 mm × 150 mm, 5 μm) and detected at 450 nm. LC-MS conditions
were set as follows: atmospheric-pressure chemical ionization (APCI) source, positive ion mode, ion source temperature of
230 ℃, charging voltage of 2 000 V and scanning range for mass-to-charge ratio of 200–1 200. The results showed that two
kinds of carotenoids were identified and their chemical formulas were C51H76O2 and C51H74O2, respectively.

Key words: carotenoid, enzymatic extraction method, high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), Rhodococcu sp. B7740

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